Hybrid cantaloupe plant bronco

ABSTRACT

A novel hybrid cantaloupe plant, designated BRONCO is disclosed. The invention relates to the seeds of cantaloupe hybrid BRONCO, to the plants and plant parts of hybrid cantaloupe BRONCO, and to methods for producing a cantaloupe plant by crossing the hybrid cantaloupe BRONCO with itself or another cantaloupe plant. The invention further relates to methods for producing a cantaloupe plant containing in its genetic material one or more transgenes and to the transgenic plants produced by that method and to methods for producing other cantaloupe plants derived from the hybrid cantaloupe plant BRONCO.

FIELD OF THE INVENTION

The present invention relates to the field of agriculture, to a new anddistinctive hybrid cantaloupe plant designated BRONCO, and to methods ofmaking and using such hybrid.

BACKGROUND OF THE INVENTION

The following description includes information that may be useful inunderstanding the present invention. It is not an admission that any ofthe information provided herein is prior art or relevant to thepresently claimed inventions, or that any publication specifically orimplicitly referenced is prior art.

Cantaloupe is an important and valuable vegetable crop. Thus, acontinuing goal of plant breeders is to develop stable, high yieldingcantaloupe hybrids that are agronomically sound. The reasons for thisgoal are to maximize the amount of fruits produced on the land used(yield) as well as to improve the plant and fruit agronomic qualities.To accomplish this goal, the cantaloupe breeder must select and developcantaloupe plants that have the traits that result in superior parentallines that combine to produce superior hybrids.

SUMMARY OF THE INVENTION

The following embodiments and aspects thereof are described inconjunction with systems, tools and methods which are meant to beexemplary, not limiting in scope.

In various embodiments, one or more of the above-described problems havebeen reduced or eliminated, while other embodiments are directed toother improvements.

According to the invention, there is provided a novel hybrid cantaloupe,designated BRONCO. This invention thus relates to the seeds of hybridcantaloupe designated BRONCO, to the plants or parts thereof of hybridcantaloupe designated BRONCO, to plants or parts thereof consistingessentially of the phenotypic and morphological characteristics ofhybrid cantaloupe designated BRONCO, and/or having all the physiologicaland morphological characteristics of hybrid cantaloupe designated BRONCOand/or having the characteristics of hybrid cantaloupe designated BRONCOlisted in Table 1 including but not limited to as determined at the 5%significance level when grown in the same environmental condition,and/or having the physiological and morphological characteristics ofhybrid cantaloupe designated BRONCO listed in Table 1 including but notlimited to as determined at the 5% significance level when grown in thesame environmental condition. The invention also relates to variants,mutants and trivial modifications of the seed or plant of hybridcantaloupe designated BRONCO.

Plant parts of the hybrid cantaloupe plant of the present invention arealso provided, such as i.e., a scion, a rootstock, a fruit, leaf,flower, cell, pollen or ovule obtained from the hybrid plant. Thepresent invention provides fruits of the hybrid cantaloupe of thepresent invention. Such fruits could be used as fresh products forconsumption or in processes resulting in processed products such asjuice, prepared fruit cuts and the like. All such products are part ofthe present invention.

The plants and seeds of the present invention include those that may beof an essentially derived variety as defined in section 41(3) of thePlant Variety Protection Act, i.e., a variety that is predominantlyderived from hybrid cantaloupe designated BRONCO or from a variety thati) is predominantly derived from hybrid cantaloupe designated BRONCO,while retaining the expression of the essential characteristics thatresult from the genotype or combination of genotypes of hybridcantaloupe designated BRONCO; ii) is clearly distinguishable from hybridcantaloupe designated BRONCO; and iii) except for differences thatresult from the act of derivation, conforms to the initial variety inthe expression of the essential characteristics that result from thegenotype or combination of genotypes of the initial variety.

In another aspect, the present invention provides regenerable cells foruse in tissue culture of hybrid cantaloupe designated BRONCO. The tissueculture will preferably be capable of regenerating plants consistingessentially of the phenotypic and morphological characteristics ofhybrid cantaloupe designated BRONCO, and/or having all the phenotypicand morphological characteristics of hybrid cantaloupe designatedBRONCO, and/or having the physiological and morphologicalcharacteristics of hybrid cantaloupe designated BRONCO, and/or havingthe characteristics of hybrid cantaloupe designated BRONCO. Preferably,the plant parts and cells used to produce such tissue cultures will beembryos, meristematic cells, seeds, callus, pollen, leaves, anthers,pistils, roots, root tips, stems, petioles, fruits, cotyledons,hypocotyls, ovaries, seed coat, fruits, endosperm, flowers or the like.Protoplasts produced from such tissue culture are also included in thepresent invention. The cantaloupe shoots, roots and whole plantsregenerated from the tissue culture, as well as the fruits produced bysaid regenerated plants are also part of the invention.

The invention also discloses methods for vegetatively propagating aplant of the present invention. Such method comprise collecting a partof a hybrid plant cantaloupe designated BRONCO and regenerating a plantfrom said part. Such part can be for example a stem cutting that isrooted into an appropriate medium according to techniques known by theone skilled in the art. Plants, part parts and fruits thereof producedby such methods are also included in the present invention. In anotheraspect, the plants and fruits thereof produced by such methods consistessentially of the phenotypic and morphological characteristics ofhybrid cantaloupe designated BRONCO, and/or having all the phenotypicand morphological characteristics of hybrid cantaloupe designatedBRONCO, and/or having the physiological and morphologicalcharacteristics of hybrid cantaloupe designated BRONCO, and/or havingthe characteristics of hybrid cantaloupe designated BRONCO.

Further included in the invention are methods for producing fruits fromthe hybrid cantaloupe designated BRONCO, said method comprising growinga hybrid cantaloupe designated BRONCO to produce a cantaloupe fruit andpreferably harvesting the hybrid cantaloupe fruit. Such fruits are partof the present invention.

Also included in this invention are methods for producing a cantaloupeplant produced by crossing the hybrid cantaloupe designated BRONCO withitself or another cantaloupe plant that can be a cantaloupe hybrid orline. When crossed with another inbred line a “three-way cross” isproduced. When crossed with itself or with another, different hybridcantaloupe, a “four-way” cross is produced. Such three and four-wayhybrid seeds and plants produced by growing said three and four-wayhybrid seeds are included in the present invention. A method forproducing a three and four-way hybrid cantaloupe seed comprisingcrossing hybrid cantaloupe designated BRONCO cantaloupe plant with adifferent cantaloupe line or hybrid and harvesting the resultant hybridcantaloupe seed are also part of the invention. The hybrid cantaloupeseed produced by the method comprising crossing hybrid cantaloupedesignated BRONCO cantaloupe plant with a different cantaloupe plant andharvesting the resultant hybrid cantaloupe seed are included in theinvention, as are included the hybrid cantaloupe plant or parts thereofand seeds produced by said grown hybrid cantaloupe plants.

Further included in the invention are methods for producing a cantaloupeseed and plants made thereof, said method comprising self-pollinatingthe hybrid cantaloupe designated BRONCO and harvesting the resultanthybrid seed. Cantaloupe seed produced by such method is also part of theinvention.

In another embodiment, this invention relates to a method for producinga hybrid cantaloupe designated BRONCO from a collection of seeds, thecollection containing both inbred line of hybrid cantaloupe designatedBRONCO seeds and hybrid seeds of BRONCO. Such a collection of seed mightbe a commercial bag of seeds. Said method comprises planting thecollection of seeds. When planted, the collection of seeds will produceinbred parent lines of hybrid cantaloupe BRONCO and hybrid plants fromthe hybrid seeds of BRONCO. Such inbred parent lines of hybridcantaloupe designated BRONCO plants are identified as having a decreasedvigor compared to the other plants grown from the collection of seeds.Said decreased vigor is due to the inbreeding depression effect and canbe identified for example by a less vigorous appearance for vegetativeand/or reproductive characteristics including a slight reduction inplant size, slightly smaller fruit size, possibly later maturity and apossible reduction in yield. If seed of the inbred lines of the hybridcantaloupe BRONCO are collected, if new inbred plants thereof are grownand crossed in a controlled manner with each other, the hybridcantaloupe BRONCO will be recreated.

This invention also relates to methods for producing other cantaloupeplants derived from hybrid cantaloupe BRONCO and to the cantaloupeplants derived by the use of those methods.

Such method for producing a cantaloupe plant derived from the hybridvariety BRONCO comprises the steps of (a) self-pollinating the hybridcantaloupe BRONCO plant at least once to produce a progeny plant; (b)crossing the progeny plant of step (a) with itself or a secondcantaloupe plant to produce a seed; (c) growing a progeny plant of asubsequent generation from the seed produced in step (b) and crossingthe progeny plant of a subsequent generation with itself or a secondcantaloupe plant to produce a cantaloupe plant derived from the hybridcantaloupe variety BRONCO. In further embodiments, steps b) or c) arerepeated for at least 1 more generation to produce a cantaloupe plantderived from the hybrid cantaloupe variety BRONCO.

Another method for producing a cantaloupe plant derived from the hybridvariety BRONCO, comprises the steps of: (a) crossing the hybridcantaloupe BRONCO plant with a second cantaloupe plant to produce aprogeny plant; (b) crossing the progeny plant of step (a) with itself ora second cantaloupe plant to produce a seed; (c) growing a progeny plantof a subsequent generation from the seed produced in step (b); (d)crossing the progeny plant of a subsequent generation of step (c) withitself or a second cantaloupe plant to produce a cantaloupe plantderived from the hybrid cantaloupe variety BRONCO. In a furtherembodiment, steps b) or c) are repeated for at least 1 more generationto produce a cantaloupe plant derived from the hybrid cantaloupe varietyBRONCO.

More specifically, the invention comprises methods for producing a malesterile cantaloupe plant, an herbicide resistant cantaloupe plant, aninsect resistant cantaloupe plant, a disease resistant cantaloupe plant,a water-stress-tolerant plant, a heat stress tolerant plants, animproved shelf life cantaloupe plant, a cantaloupe plant with increasedsweetness and flavor, a cantaloupe plant with increased sugar content, acantaloupe plant with delayed senescence or controlled ripening orplants with improved salt tolerance. Said methods comprise transformingthe hybrid designated BRONCO cantaloupe plant with nucleic acidmolecules that confer male sterility, herbicide resistance, insectresistance, disease resistance, water-stress tolerance, heat stresstolerance, increased shelf life, increased sweetness and flavor,increased sugar content, delayed senescence or controlled ripening orimproved salt tolerance, respectively. The transformed cantaloupe plantsor parts thereof, obtained from the provided methods, including forexample a male sterile cantaloupe plant, an herbicide resistantcantaloupe plant, an insect resistant cantaloupe plant, a diseaseresistant cantaloupe plant, a cantaloupe with water stress tolerance, acantaloupe with heat stress tolerance, a cantaloupe plants withincreased sweetness and flavor, a cantaloupe plants with increased sugarcontent, a cantaloupe plants with delayed senescence or controlledripening or a cantaloupe plants with improved salt tolerance areincluded in the present invention. Plants may display one or more of theabove listed traits. For the present invention and the skilled artisan,disease is understood to be fungal diseases, viral diseases, bacterialdiseases, mycoplasm diseases, or other plant pathogenic diseases and adisease resistant plant will encompass a plant resistant to fungal,viral, bacterial, mycoplasm, and other plant pathogens.

In another aspect, the present invention provides for methods ofintroducing one or more desired trait(s) into the hybrid cantaloupeBRONCO and plants or seeds obtained from such methods. The desiredtrait(s) may be, but not exclusively, a single gene, preferably adominant but also a recessive allele. Preferably, the transferred geneor genes will confer such traits as male sterility, herbicideresistance, insect resistance, resistance for bacterial, fungal,mycoplasma or viral disease, improved shelf life, water-stresstolerance, delayed senescence or controlled ripening, enhancednutritional quality such as increased sugar content or increasedsweetness, enhanced plant quality such as improved drought or salttolerance, enhanced plant vigor or improve fresh cut application,specific aromatic compounds, specific volatiles, flesh texture; specificnutritional components and long shelf life (LSL). The gene or genes maybe naturally occurring cantaloupe gene(s), mutant(s) or transgene(s)introduced through genetic engineering techniques. The method forintroducing the desired trait(s) is preferably a backcrossing processmaking use of a series of backcrosses to at least one of the parentlines of hybrid cantaloupe BRONCO during which the desired trait(s) ismaintained by selection. The single gene conversion plants that can beobtained by the method are included in the present invention

When using a transgene, the trait is generally not incorporated intoeach newly developed hybrid such as BRONCO by direct transformation.Rather, the more typical method used by breeders of ordinary skill inthe art to incorporate the transgene is to take a line already carryingthe transgene and to use such line as a donor line to transfer thetransgene into one or more of the parents of the newly developed hybrid.The same would apply for a naturally occurring trait or one arising fromspontaneous or induced mutations. The backcross breeding processcomprises the following steps: (a) crossing one of the parental inbredline plants of BRONCO with plants of another line that comprise thedesired trait(s), (b) selecting the F1 progeny plants that have thedesired trait(s); (c) crossing the selected F1 progeny plants with theparental inbred cantaloupe lines of hybrid BRONCO plants to producebackcross progeny plants; (d) selecting for backcross progeny plantsthat have the desired trait(s) and physiological and morphologicalcharacteristics of the cantaloupe parental inbred line of hybridcantaloupe BRONCO to produce selected backcross progeny plants; and (e)repeating steps (c) and (d) one, two, three, four, five six, seven,eight, nine or more times in succession to produce selected, second,third, fourth, fifth, sixth, seventh, eighth, ninth or higher backcrossprogeny plants that consist essentially of the phenotypic andmorphological characteristics of the parental inbred cantaloupe line ofhybrid cantaloupe BRONCO, and/or have all the phenotypic andmorphological characteristics of the parental cantaloupe inbred line ofhybrid cantaloupe BRONCO, and/or have the desired trait(s) and thephysiological and morphological characteristics of the parental inbredcantaloupe line of cantaloupe hybrid BRONCO as determined in Table 1,including but not limited to at a 5% significance level when grown inthe same environmental conditions. The cantaloupe plants or seedproduced by the methods are also part of the invention, as are thehybrid cantaloupe BRONCO plants that comprised the desired trait.Backcrossing breeding methods, well known to one skilled in the art ofplant breeding will be further developed in subsequent parts of thespecification.

In an embodiment of this invention is a method of making a backcrossconversion of hybrid cantaloupe BRONCO, comprising the steps of crossingone of the parental cantaloupe inbred line plants of hybrid BRONCO witha donor plant comprising a mutant gene(s), a naturally occurringgene(s), or transgene(s) conferring one or more desired trait, selectingan F1 progeny plant comprising the naturally occurring gene(s), mutantgene(s) or transgene(s) conferring the one or more desired trait, andbackcrossing the selected F1 progeny plant to the parental cantaloupeinbred line plants of hybrid BRONCO. This method may further comprisethe step of obtaining a molecular marker profile of the parentalcantaloupe inbred line plants of hybrid BRONCO and using the molecularmarker profile to select for a progeny plant with the desired trait andthe molecular marker profile of the parental cantaloupe inbred lineplants of hybrid BRONCO. This method further comprises crossing theparental inbred cantaloupe line plants of hybrid cantaloupe BRONCOcontaining the naturally occurring gene(s), the mutant gene(s) or thetransgene(s) conferring the one or more desired trait with the secondparental inbred cantaloupe line plants of hybrid cantaloupe BRONCO inorder to produce the hybrid cantaloupe BRONCO comprising the naturallyoccurring gene(s), the mutant gene(s) or transgene(s) conferring the oneor more desired trait. The plants or parts thereof produced by suchmethods are also part of the present invention.

In some embodiments of the invention, the number of loci that may bebackcrossed into the parental cantaloupe inbred line of hybrid BRONCO isat least 1, 2, 3, 4, or 5. A single locus may contain severaltransgenes, such as a transgene for disease resistance that, in the sameexpression vector, also contains a transgene for herbicide resistance.The gene for herbicide resistance may be used as a selectable markerand/or as a phenotypic trait. A single locus conversion of site specificintegration system allows for the integration of multiple genes at theconverted locus.

The invention further provides methods for developing cantaloupe plantsin a cantaloupe plant breeding program using plant breeding techniquesincluding recurrent selection, backcrossing, pedigree breeding,molecular marker (Isozyme Electrophoresis, Restriction Fragment LengthPolymorphisms (RFLPs), Randomly Amplified Polymorphic DNAs (RAPDs),Arbitrarily Primed Polymerase Chain Reaction (AP-PCR), DNA AmplificationFingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs),Amplified Fragment Length Polymorphisms (AFLPs), and Simple SequenceRepeats (SSRs) which are also referred to as Microsatellites, etc.)enhanced selection, genetic marker enhanced selection andtransformation. Seeds, cantaloupe plants, and parts thereof produced bysuch breeding methods are also part of the invention.

The invention also relates to variants, mutants and trivialmodifications of the seed or plant of the cantaloupe hybrid BRONCO orinbred parental lines thereof. Variants, mutants and trivialmodifications of the seed or plant of hybrid cantaloupe BRONCO or inbredparental lines thereof can be generated by methods available to oneskilled in the art, including but not limited to, mutagenesis (e.g.,chemical mutagenesis, radiation mutagenesis, transposon mutagenesis,insertional mutagenesis, signature tagged mutagenesis, site-directedmutagenesis, and natural mutagenesis), knock-outs/knock-ins, antisenseand RNA interference. For more information of mutagenesis in plants,such as agents, protocols, see Acquaah et al. (Principles of plantgenetics and breeding, Wiley-Blackwell, 2007, ISBN 1405136464,9781405136464, which is herein incorporated by reference in its entity).

The invention also relates to a mutagenized population of the hybridcantaloupe BRONCO and methods of using such populations. In someembodiments, the mutagenized population can be used in screening for newcantaloupe plants which comprises one or more or all of themorphological and physiological characteristics of hybrid cantaloupeBRONCO. In some embodiments, the new cantaloupe plants obtained from thescreening process comprise all of the morphological and physiologicalcharacteristics of the cantaloupe hybrid BRONCO, and one or moreadditional or different morphological and physiological characteristicsthat the cantaloupe hybrid BRONCO does not have.

This invention also is directed to methods for producing a cantaloupeplant by crossing a first parent cantaloupe plant with a second parentcantaloupe plant wherein either the first or second parent cantaloupeplant is a hybrid cantaloupe plant of BRONCO. Further, both first andsecond parent cantaloupe plants can come from the hybrid cantaloupeplant BRONCO. Further, the hybrid cantaloupe plant BRONCO can beself-pollinated i.e. the pollen of a hybrid cantaloupe plant BRONCO canpollinate the ovule of the same hybrid cantaloupe plant BRONCO. Whencrossed with another cantaloupe plant, a hybrid seed is produced. Suchmethods of hybridization and self-pollination are well known to thoseskilled in the art of breeding.

An inbred cantaloupe line such as one of the parental lines of hybridcantaloupe BRONCO has been produced through several cycles ofself-pollination and is therefore to be considered as a homozygous line.An inbred line can also be produced though the dihaploid system whichinvolves doubling the chromosomes from a haploid plant or embryo thusresulting in an inbred line that is genetically stable (homozygous) andcan be reproduced without altering the inbred line: Haploid plants couldbe obtained from haploid embryos that might be produced frommicrospores, pollen, anther cultures or ovary cultures. The haploidembryos may then be doubled by chemical treatments such as by colchicineor be doubled autonomously. The haploid embryos may also be grown intohaploid plants and treated to induce the chromosome doubling. In eithercase, fertile homozygous plants are obtained. A hybrid variety isclassically created through the fertilization of an ovule from an inbredparental line by the pollen of another, different inbred parental line.Due to the homozygous state of the inbred line, the produced gametescarry a copy of each parental chromosome. As both the ovule and thepollen bring a copy of the arrangement and organization of the genespresent in the parental lines, the genome of each parental line ispresent in the resulting F1 hybrid, theoretically in the arrangement andorganization created by the plant breeder in the original parental line.

As long as the homozygosity of the parental lines is maintained, theresulting hybrid cross shall be stable. The F1 hybrid is then acombination of phenotypic characteristics issued from two arrangementand organization of genes, both created by a man skilled in the artthrough the breeding process.

Still further, this invention also is directed to methods for producinga hybrid cantaloupe plant BRONCO-derived cantaloupe plant by crossinghybrid cantaloupe plant BRONCO with a second cantaloupe plant andgrowing the progeny seed, and possibly repeating the crossing andgrowing steps with the hybrid cantaloupe plant BRONCO-derived plant from0 to 7 times. Thus, any such methods using the hybrid cantaloupe plantBRONCO are part of this invention: selfing, backcrosses, hybridproduction, crosses to populations, and the like. All plants producedusing hybrid cantaloupe plant BRONCO as a parent are within the scope ofthis invention, including plants derived from hybrid cantaloupe plantBRONCO. Such plants might exhibit additional and desired characteristicsor traits such as high seed yield, high seed germination, seedlingvigor, early fruit maturity, high fruit yield, ease of fruit setting,disease tolerance or resistance, and adaptability for soil and climateconditions. Consumer-driven traits, such as a preference for a givenfruit size, shape, color, texture, and taste are other traits that maybe incorporated into new cantaloupe plants developed by this invention.

A cantaloupe plant can also be propagated vegetatively. A part of theplant, for example a shoot tissue, is collected, and a new plant isobtained from the part. Such part typically comprises an apical meristemof the plant. The collected part is transferred to a medium allowingdevelopment of a plantlet, including for example rooting or developmentof shoots, or is grafted onto a cantaloupe plant or a rootstock preparedto support growth of shoot tissue. This is achieved using methodswell-known in the art. Accordingly, in one embodiment, a method ofvegetatively propagating a plant of the present invention comprisescollecting a part of a plant according to the present invention, e.g. ashoot tissue, and obtaining a plantlet from said part. In oneembodiment, a method of vegetatively propagating a plant of the presentinvention comprises: a) collecting tissue of a plant of the presentinvention; b) rooting said proliferated shoots to obtain rootedplantlets. In one embodiment, a method of vegetatively propagating aplant of the present invention comprises: a) collecting tissue of aplant of the present invention; b) cultivating said tissue to obtainproliferated shoots; c) rooting said proliferated shoots to obtainrooted plantlets. In one embodiment, such method further comprisesgrowing a plant from said plantlets. In one embodiment, a fruit isharvested from said plant. In one embodiment, the fruit is processedinto products such as canned cantaloupe, juice, fresh cut fruits and thelike.

In some embodiments, the present invention teaches a seed of hybridcantaloupe designated BRONCO, wherein a representative sample of seed ofsaid hybrid is deposited under NCIMB No 42565.

In some embodiments, the present invention teaches a cantaloupe plant,or a part thereof, produced by growing the deposited BRONCO seed.

In some embodiments, the present invention teaches cantaloupe plantparts, wherein the cantaloupe part is selected from the group consistingof: a leaf, a flower, a fruit, an ovule, pollen, a cell a rootstock, anda scion.

In some embodiments, the present invention teaches a cantaloupe plant,or a part thereof, having all of the characteristics of hybrid BRONCO aslisted in Table 1 of this application.

In some embodiments, the present invention teaches a cantaloupe plant,or a part thereof, having all of the physiological and morphologicalcharacteristics of hybrid BRONCO, wherein a representative sample ofseed of said hybrid was deposited under NCIMB No 42565.

In some embodiments, the present invention teaches a tissue culture ofregenerable cells produced from the plant or plant part grown from thedeposited BRONCO seed, wherein cells of the tissue culture are producedfrom a plant part selected from the group consisting of protoplasts,embryos, meristematic cells, callus, pollen, ovules, flowers, seeds,leaves, roots, root tips, anthers, stems, petioles, fruits, cotyledonsand hypocotyls. In some embodiments, the plant part includes protoplastsproduced from a plant grown from the deposited BRONCO seed.

In some embodiments, the present invention teaches a cantaloupe plantregenerated from the tissue culture from a plant grown from thedeposited BRONCO seed, said plant having the characteristics of hybridBRONCO, wherein a representative sample of seed of said hybrid isdeposited under NCIMB No 42565.

In some embodiments, the present invention teaches a cantaloupe fruitproduced from the plant grown from the deposited BRONCO seed.

In some embodiments, methods of producing said cantaloupe fruit comprisea) growing the cantaloupe plant from deposited BRONCO seed to produce acantaloupe fruit, and b) harvesting said cantaloupe fruit. In someembodiments, the present invention also teaches a cantaloupe fruitproduced by the method of producing cantaloupe fruit as described above.

In some embodiments, the present invention teaches methods for producinga cantaloupe seed comprising crossing a first parent cantaloupe plantwith a second parent cantaloupe plant and harvesting the resultantcantaloupe seed, wherein said first parent cantaloupe plant and/orsecond parent cantaloupe plant is the cantaloupe plant produced from thedeposited BRONCO seed.

In some embodiments, the present invention teaches methods for producinga cantaloupe seed comprising self-pollinating the cantaloupe plant grownfrom the deposited BRONCO seed and harvesting the resultant cantaloupeseed.

In some embodiments, the present invention teaches the seed produced byany of the above described methods.

In some embodiments, the present invention teaches methods ofvegetatively propagating the cantaloupe plant grown from the depositedBRONCO seed, said method comprising a) collecting part of a plant grownfrom the deposited BRONCO seed and b) regenerating a plant from saidpart.

In some embodiments, the method further comprises harvesting a fruitfrom said vegetatively propagated plant.

In some embodiments, the present invention teaches the plant and thefruit of plants vegetatively propagated from plant parts of plants grownfrom the deposited BRONCO seed.

In some embodiments, the present invention teaches methods of producinga cantaloupe plant derived from the hybrid variety BRONCO, the methodcomprising the steps of: (a) self-pollinating the plant of grown fromthe deposited BRONCO seed at least once to produce a progeny plant; (b)crossing the progeny plant of step (a) with itself or a secondcantaloupe plant to produce a seed; (c) growing a progeny plant of asubsequent generation from the seed produced in step (b), and crossingthe progeny plant of a subsequent generation with itself or a secondcantaloupe plant to produce a cantaloupe plant derived from the hybridcantaloupe variety BRONCO. In some embodiments said method furthercomprises the step of: (d) repeating steps b) or c) for at least 1 moregeneration to produce a cantaloupe plant derived from the hybridcantaloupe variety BRONCO.

In some embodiments, the present invention teaches methods of producinga cantaloupe plant derived from the hybrid variety BRONCO, the methodscomprising the steps of: (a) crossing the plant grown from the depositedBRONCO seed with a second cantaloupe plant to produce a progeny plant;(b) crossing the progeny plant of step (a) with itself or a secondcantaloupe plant to produce a seed; (c) growing a progeny plant of asubsequent generation from the seed produced in step (b); (d) crossingthe progeny plant of a subsequent generation of step (c) with itself ora second cantaloupe plant to produce a cantaloupe plant derived from thehybrid cantaloupe variety BRONCO. In some embodiments said methodfurther comprises the steps of: (d) repeating step b) or c) for at least1 more generation to produce a cantaloupe plant derived from the hybridcantaloupe variety BRONCO.

In some embodiments, the present invention teaches methods for producinga transgenic cantaloupe plant, the methods comprising crossing a firstcantaloupe plant grown from the deposited BRONCO seed with a secondcantaloupe plant containing a transgene, wherein the transgene of saidsecond cantaloupe plant is integrated into the genome of the cantaloupeplant progeny resulting from said cross, and wherein the transgeneconfers said cantaloupe plant progeny with at least one trait selectedfrom the group consisting of male sterility, male fertility, herbicideresistance, insect resistance, disease resistance, increased sweetness,increased sugar content, increased flavor, improved ripening control,and improved salt tolerance.

In some embodiments, the present invention teaches methods for producinga transgenic cantaloupe plant, the method comprising transforming atleast one transgene into a hybrid cantaloupe BRONCO plant, or a plantpart or a plant cell thereof or parental line used for producing thehybrid cantaloupe plant BRONCO, a sample seed of said hybrid having beendeposited under NCIMB Accession No. 42565, thereby producing atransgenic cantaloupe plant.

In some embodiments, the present invention teaches cantaloupe plants andcantaloupe fruits produced by any of the above-described methods ofproducing transgenic cantaloupes. Thus, in some embodiments, the presentinvention teaches a plant grown from the deposited BRONCO seed, furthercomprising a transgene. In some embodiments, said transgenic cantaloupecomprise transgenes which confer said plant with a trait selected fromthe group consisting of male sterility, male fertility, herbicideresistance, insect resistance, disease resistance, increased sweetness,increased sugar content, increased flavor, improved ripening control,and improved salt tolerance.

In some embodiments, the present invention teaches plants grown from thedeposited BRONCO seed wherein said plants comprise at least one singlelocus conversion. In some embodiments said single locus conversionconfers said plant with a trait selected from the group consisting ofmale sterility, male fertility, herbicide resistance, insect resistance,disease resistance, increased sweetness, increased sugar content,increased flavor, improved ripening control, long shelf life, specificaromatic compounds and improved salt tolerance. In some embodiments, theat least one single locus conversion is an artificially mutated gene.

In addition to the exemplary aspects and embodiments described above,further aspects and embodiments will become apparent by study of thefollowing descriptions.

DETAILED DESCRIPTION OF THE INVENTION

Definitions

In the description and tables that follow, a number of terms are used.In order to provide a clear and consistent understanding of thespecification and claims, including the scope to be given such terms,the following definitions are provided:

Abscission zone. This is the zone of abscission or separation of thefruit from the peduncle at maturity (controlled by ethylene). Theresulting zone (or scar) ranges in size, small being preferred overlarge—range small (<10 mm), medium (10-15 mm), large (15-20 mm), verylarge (>20 mm)

Allele. An allele is any of one or more alternative forms of a genewhich relate to one trait or characteristic. In a diploid cell ororganism, the two alleles of a given gene occupy corresponding loci on apair of homologous chromosomes.

Backcrossing. Backcrossing is a process in which a breeder repeatedlycrosses hybrid progeny back to one of the parents, for example, a firstgeneration hybrid F₁ with one of the parental genotype of the F₁ hybrid.

Blossom scar. This is the remnant scar from the stigmatic surface of theblossom. There is a very broad range in sizes, small is better. Range issmall (<10 mm), medium (10-20 mm), large (20-40 mm) and very large (>40mm).

Cavity. As used herein, cavity refers to the center of the cantaloupefruit containing seeds and maternal tissues. Cavity measurements aremade on a single fruit or recorded as an average of many fruit atharvest maturity and recorded in a convenient unit of measure. Cavityratings: 1=very poor (non marketable), 3=poor (non marketable),5=average (marketable) 7=very good (much better than industrystandards), 9=superior (further improvement not attainable).

Cavity to overall fruit diameter, ratio. Cavity to Diameter ratio is ameasure of the cavity size compared to the overall fruit size of asingle fruit or the average of many fruit at harvest maturity andrecorded in a convenient unit of measure. In some embodiments lowerratios of fruit cavity to overall fruit diameter are desired.

Collection of seeds. In the context of the present invention acollection of seeds is a grouping of seeds mainly containing similarkind of seeds, for example hybrid seeds having the inbred line of theinvention as a parental line, but that may also contain, mixed togetherwith this first kind of seeds, a second, different kind of seeds, of oneof the inbred parent lines, for example the inbred line of the presentinvention. A commercial bag of hybrid seeds having the inbred line ofthe invention as a parental line and containing also the inbred lineseeds of the invention would be, for example such a collection of seeds.

Essentially all the physiological and morphological characteristics. Aplant having essentially all the physiological and morphologicalcharacteristics means a plant having the physiological and morphologicalcharacteristics of the recurrent parent, except for the characteristicsderived from the converted gene.

Grafting. Grafting is the operation by which a rootstock is grafted witha scion. The primary motive for grafting is to avoid damages bysoil-born pest and pathogens when genetic or chemical approaches fordisease management are not available. Grafting a susceptible scion ontoa resistant rootstock can provide a resistant cultivar without the needto breed the resistance into the cultivar. In addition, grafting mayenhance tolerance to abiotic stress, increase yield and result in moreefficient water and nutrient uses.

Immunity to disease(s) and or insect(s). A cantaloupe plant which is notsubject to attack or infection by specific disease(s) and or insect(s)is considered immune.

Intermediate/Moderate resistance to disease(s) and or insect(s). Acantaloupe plant that restricts the growth and development of specificdisease(s) and or insect(s), but may exhibit a greater range of symptomsor damage compared to high/standard resistant plants. Intermediateresistant plants will usually show less severe symptoms or damage thansusceptible plant varieties when grown under similar environmentalconditions and/or specific disease(s) and or insect(s) pressure, but mayhave heavy damage under heavy pressure. Intermediate resistant tomatoplants are not immune to the disease(s) and or insect(s).

Long shelf life (LSL). Long shelf life melons are melon where theinhibition of a normal ripening pattern results in delayed maturity, butincreased field holding and harvest flexibility.

Monecious. The term used to describe a plant variety where each flowerexhibits only one sexual character (either male or female) and eachplant has flowers of both sexes.

Netting. The height and density of the netting (reticulation) thatcovers orange flesh melons. Range is fine, medium, medium coarse andcoarse. (i.e.—a fine net would be low and would have noticeable spacebetween the net, a coarse net would be quite high and almost completelycover the fruit exterior. Ideal net is medium or medium coarse. Nettingcan also be assigned a descriptive number 1=fine net to 10 coarse net.

Number of Boxes per Acre. The Number of Boxes per Acre—6's, 9's, 12's,15's, 18's or 23's refers to the number of fruit that fit into astandard cantaloupe box.

Overall Rating. A final or Overall Rating is assigned to varietyperformance or a varieties characteristic in test or trial situations ofa variety. Overall Rating can range from 1=very poor to 10 excellent.

Oval. Oval is used to describe fruit shape when the length is greaterthan the width and ranges from a slight oval, oval to heavy oval.

Plant Cell. Plant cell, as used herein includes plant cells whetherisolated, in tissue culture or incorporated in a plant or plant part.

Plant Part. As used herein, the term plant includes plant cells, plantprotoplasts, plant cell tissue cultures from which cantaloupe plants canbe regenerated, plant calli, plant clumps and plant cells that areintact in plants or parts of plants, such as embryos, pollen, ovules,flowers, seeds, fruit, rootstock, scions, stems, roots, anthers,pistils, root tips, leaves, meristematic cells, axillary buds,hypocotyls cotyledons, ovaries, seed coat endosperm and the like.

Quantitative Trait Loci (QTL) Quantitative trait loci refer to geneticloci that control to some degree numerically representable traits thatare usually continuously distributed.

Regeneration. Regeneration refers to the development of a plant fromtissue culture.

Resistance to disease(s) and or insect(s). A cantaloupe plant thatrestricts highly the growth and development of specific disease(s) andor insect(s) under normal disease(s) and or insect(s) attack pressurewhen compared to susceptible plants. These cantaloupe plants can exhibitsome symptoms or damage under heavy disease(s) and or insect(s)pressure. Resistant cantaloupe plants are not immune to the disease(s)and or insect(s).

Rind Contrast. A subjective measure of the color difference between therind and the fruit flesh. 1=no contrast to 10=excellent contrast.

Rootstock. A rootstock is the lower part of a plant capable of receivinga scion in a grafting process.

Season maturity or maturity. Maturity is considered the date of theonset of harvest and is classified as Very Early, Early, Mid Early, Mainand Late or specified by recording the date of the onset of harvest. Insome embodiments, under summer conditions in Davis Calif., very earlymaturity is reached within 72-76 days, early maturity is within 76-80days, mid early maturity is within 80-84 days, main maturity is between84-88 days, and late maturity is between 88-92 days.

Scion: A scion is the higher part of a plant capable of being graftedonto a rootstock in a grafting process.

Single gene converted (conversion). Single gene converted (conversion)plants refer to plants which are developed by a plant breeding techniquecalled backcrossing wherein essentially all of the desired morphologicaland physiological characteristics of a plant are recovered in additionto the single gene transferred into the plant via the backcrossingtechnique or via genetic engineering.

Susceptible to disease(s) and or insect(s). A cantaloupe plant that issusceptible to disease(s) and or insect(s) is defined as a cantaloupeplant that has the inability to restrict the growth and development ofspecific disease(s) and or insect(s). Plants that are susceptible willshow damage when infected and are more likely to have heavy damage undermoderate levels of specific disease(s) and or insect(s).

Shape. Refers to external fruit shape. Range is Flat Round, Round, roundoval, oval, elongate.

Soluble Solids. Soluble solids refer to the percent of solid materialfound in the fruit tissue, the vast majority of which is sugars. Solublesolids are estimated with a refractometer and measured as degrees Brix.Soluble Solids vary with environment. For example, for California summergrowing conditions the following range would apply. Very high (>12.5%),high (11.5-12.5%), medium (10.5-11.5%), low <10.5%).

Tolerance to abiotic stresses. A cantaloupe plant that is tolerant toabiotic stresses has the ability to endure abiotic stress withoutserious consequences for growth, appearance and yield.

Uniformity. Uniformity, as used herein, describes the similarity betweenplants or plant characteristics which can be a described by qualitativeor quantitative measurements.

Variety. A plant variety as used by one skilled in the art of plantbreeding means a plant grouping within a single botanical taxon of thelowest known rank which can be defined by the expression of thecharacteristics resulting from a given genotype or combination ofphenotypes, distinguished from any other plant grouping by theexpression of at least one of the said characteristics and considered asa unit with regard to its suitability for being propagated unchanged(International convention for the protection of new varieties ofplants).

Vine Overall. An overall rating assigned to the performance of a plant'svine. Vine Overall can range from 1=very poor to 10 excellent.

Yield. Yield Type is defined as concentrated, semi concentrated orextended. Concentrated=harvested yield produced in consecutive (4-5days) of harvest. Semi concentrated=harvested yield produced within 7-9days. Extended=harvested yield is produced over the course of 10-14days. The Fruit Set may also be defined accordingly to the samecriteria, i.e. very concentrated, when the plant sets all of its fruitat nearly the same time; concentrated, when the plant sets all itsfruits in a short period of time; semi concentrated, when fruit set isless uniform; and extended, when the plant sets and matures fruit toallow picking over a long period of time.

Yield Rating is defined an overall rating assigned to yield. YieldRating can range from 1=very poor to 9 excellent.

Cantaloupe Plants

Practically speaking, all cultivated forms of cantaloupe belong to thehighly polymorphic species Cucumis melo L. that is grown for its sweetedible fruit. The term cantaloupe, as used herein, refers to theAmerican usage of the term which is used to describe the netted melonscommonly referred to as cantaloupe or muskmelon in U.S. commerce. As acrop, cantaloupes are grown commercially wherever environmentalconditions permit the production of an economically viable yield. Theyare produced on non-climbing vines that are cultivated prostrate on thesoil. On healthy plants there is a canopy of large, soft, hairy leaves,generally heart shaped and somewhat lobed. Fruits may be orange fleshedor green fleshed.

The fruit surface is generally netted and roughened and in somevarieties sutured. Fruit shape is generally round to oval and ranges insize from five to eight inches long and about equal in diameter.Cantaloupe is considered a very tender warm season crop but can beproduced in all areas of the World where optimum temperatures of 65-75 Foccur for at least 120 frost-free days. Commercial yields are considered“good” when over 20,000 of marketable fruit are harvested per acre.Typically, productions fields are harvested multiple times by hand andthe fruit may be picked before ripening and shipped long distances tothe end market. In the United States, the principal fresh marketcantaloupe growing regions are California, Arizona and Texas whichproduce approximately 96,000 acres out of a total annual acreage of morethan 113,000 acres (USDA, 1998).

Fresh cantaloupes are available in the United States year-round althoughthe greatest supply is from June through October. Fresh cantaloupes areconsumed in many forms. They are eaten sliced or diced and used as aningredient in many prepared foods, such as the popular consumption ofmelon pieces wrapped in prosciutto ham.

Cucumis melo is a member of the family Cucurbitaceae. The Cucurbitaceaeis a family of about 90 genera and 700 to 760 species, mostly of thetropics. The family includes pumpkins, squashes, gourds, watermelon,loofah and several weeds. The genus Cucumis, to which the cantaloupe,cucumbers, and several melons belong, includes about 70 species. Cucumismelo includes a wide range of cultivated plants. Although crossesoutside the species are sterile, intraspecific crosses are generallyfertile, resulting in a confusing range of variation. The more commoncultivated plants fall into four main groups. First are the truecantaloupes of Europe. These have thick, scaly, rough, often deeplygrooved, but not netted rinds. Second are the muskmelons, mostly grownin the United States, where they are incorrectly called cantaloupes.These have finely netted rinds with shallow ribs. Third are the casabaor winter melons with large fruits. These have smooth, often yellowrinds. The honeydew melons are in this third group. Fourth are a groupof elongated melons of India, China and Japan which are grown asvegetables. Other classification schemes and cultivars could bepresented.

Cantaloupe is a simple diploid species with twelve pairs of highlydifferentiated chromosomes. The Cucumis melo genome includes over 375 Mbof sequence with an estimated 27,427 protein-coding genes (Garcia-Mas etal., (2012. The genome of melon (Cucumis melo L.). PNAS July issue).

Large field spaces are required for cantaloupe and the need for laborintensive hand pollination for self as well as cross pollination hasresulted in a lag in the knowledge of cantaloupe genetics relative tosuch crops as tomato. Cantaloupe flowers open after sunrise; the exacttime depends on environmental conditions such as sunlight, temperatureand humidity. The flower closes permanently in the afternoon of the sameday. Almost all pollen is collected and transferred before noon.Typically flowers are staminate although some are also hermaphroditic.Although hermaphroditic flowers are self-fertile, they are incapable ofperforming self-pollination. Insects are required for pollination. Theprimary pollinators are bees, particularly honey bees.

There are numerous steps in the development of any novel, desirableplant germplasm. Plant breeding begins with the analysis and definitionof problems and weaknesses of the current germplasm, the establishmentof program goals, and the definition of specific breeding objectives.The next step is selection of germplasm that possesses the traits tomeet the program goals. The goal is to combine in a single variety orhybrid an improved combination of desirable traits from the parentalgermplasm.

In cantaloupe, these important traits may include higher yield, fieldperformance, fruit and agronomic quality such as sugar levels, smallcavity size, flesh color or texture, rind firmness or strong net,resistance to diseases and insects, ease of fruit setting, adaptabilityfor soil and climate conditions, field holding, harvest flexibility andtolerance to drought and heat.

Particularly desirable traits that may be incorporated by this inventionare improved resistance to different viral, fungal, and bacterialpathogens and improved resistance to insect pests. Important diseasesinclude but are not limited to Powdery mildew (Sphaerothica fuligenea),Fusarium wilt race 0,1,2 (Fusarium oxysporum melonis), Downy mildew(Pseudoperonospora cubensis), Cucumber mosaic virus, watermelon mosaicvirus, zucchini mosaic virus, papaya ringspot virus, melon aphid (aphysgossyppi).

Other desirable traits include traits related to improved cantaloupefruits. A non-limiting list of fruit phenotypes used during breedingselection include:

-   -   Firm fruit exterior. Fruit Firmness subjectively tested under        field conditions for resistance of fruit exterior against a        given pressure. Range is soft, medium, firm and very firm.    -   Flesh color. Flesh color is defined as degree of intensity of        orange. Flesh color ratings 1=very poor (non marketable), 3=poor        (non marketable), 5=average (marketable) 7=very good (much        better than industry standards), 9=superior (further improvement        not attainable).    -   Flesh firmness. Flesh firmness subjectively tested under field        conditions for resistance of flesh against a given pressure.        Firmness ratings 1=very poor (non marketable), 3=poor (non        marketable), 5=average (marketable) 7=very good (much better        than industry standards), 9=superior (further improvement not        attainable).    -   Fruit Diameter. The cross-sectional diameter of a single fruit        of the average of many fruit measured at harvest maturity and        recorded in a convenient unit of measure.    -   Fruit Length. The longitudinal length of a single fruit or the        average of many fruit measure from stem to blossom end at        harvest maturity and recorded in a convenient unit of measure.    -   Fruit size. Western Shipper fruit size determined two ways 1/.        Range in kilograms: small (below 1.5), medium (1.5-1.8), large        (1.8-2.2), very large (above 2.2) 2/. # Fruit that fit into a        standard western melon packing box: 6, 9, 12, 15, 18, 23, 30.        Small: some 18's, 23's, 30's, Medium: some 12's, 15's 18's,        Large: 9's, 12's, few 15's and Extra Large: few 6's, 9's few        12's.    -   Fruit Weight. The weight of a single fruit or the average of        many fruit measured at harvest maturity and recorded in a        convenient unit of measure.        Cantaloupe Breeding

The goal of cantaloupe breeding is to develop new, unique and superiorcantaloupe inbred lines and hybrids. The breeder initially selects andcrosses two or more parental lines, followed by repeated selfing andselection, producing many new genetic combinations. Another method usedto develop new, unique and superior cantaloupe inbred lines and hybridsoccurs when the breeder selects and crosses two or more parental linesfollowed by haploid induction and chromosome doubling that result in thedevelopment of dihaploid inbred lines. The breeder can theoreticallygenerate billions of different genetic combinations via crossing,selfing and mutations and the same is true for the utilization of thedihaploid breeding method.

Each year, the plant breeder selects the germplasm to advance to thenext generation. This germplasm is grown under unique and differentgeographical, climatic and soil conditions, and further selections arethen made, during and at the end of the growing season. The inbred linesdeveloped are unpredictable. This unpredictability is because thebreeder's selection occurs in unique environments, with no control atthe DNA level (using conventional breeding procedures or dihaploidbreeding procedures), and with millions of different possible geneticcombinations being generated. A breeder of ordinary skill in the artcannot predict the final resulting lines he develops, except possibly ina very gross and general fashion. This unpredictability results in theexpenditure of large research monies to develop superior new cantaloupeinbred lines and hybrids.

The development of commercial cantaloupe hybrids requires thedevelopment of homozygous inbred lines, the crossing of these lines, andthe evaluation of the hybrid crosses.

Pedigree breeding and recurrent selection breeding methods are used todevelop inbred lines from breeding populations. Breeding programscombine desirable traits from two or more inbred lines or variousbroad-based sources into breeding pools from which inbred lines aredeveloped by selfing and selection of desired phenotypes or through thedihaploid breeding method followed by the selection of desiredphenotypes. The new inbreds are crossed with other inbred lines and thehybrids from these crosses are evaluated to determine which havecommercial potential.

Choice of breeding or selection methods depends on the mode of plantreproduction, the heritability of the trait(s) being improved, and thetype of cultivar used commercially (e.g., F₁ hybrid cultivar, purelinecultivar, etc.). For highly heritable traits, a choice of superiorindividual plants evaluated at a single location will be effective,whereas for traits with low heritability, selection should be based onmean values obtained from replicated evaluations of families of relatedplants. Popular selection methods commonly include pedigree selection,modified pedigree selection, mass selection, recurrent selection, andbackcross breeding.

i Pedigree Selection

Pedigree breeding is used commonly for the improvement ofself-pollinating crops or inbred lines of cross-pollinating crops. Twoparents possessing favorable, complementary traits are crossed toproduce an F₁. An F₂ population is produced by selfing one or severalF₁s or by intercrossing two F₁s (sib mating). The dihaploid breedingmethod could also be used. Selection of the best individuals is usuallybegun in the F₂ population; then, beginning in the F₃, the bestindividuals in the best families are selected. Replicated testing offamilies, or hybrid combinations involving individuals of thesefamilies, often follows in the F₄ generation to improve theeffectiveness of selection for traits with low heritability. At anadvanced stage of inbreeding (i.e., F₆ and F₇), the best lines ormixtures of phenotypically similar lines are tested for potential use asparents of new hybrid cultivars. Similarly, the development of newinbred lines through the dihaploid system requires the selection of thebest inbreds followed by two to five years of testing in hybridcombinations in replicated plots.

ii Backcross Breeding

Backcross breeding has been used to transfer genes for a simplyinherited, highly heritable trait into a desirable homozygous cultivaror inbred line which is the recurrent parent. The source of the trait tobe transferred is called the donor parent. The resulting plant isexpected to have the attributes of the recurrent parent (e.g., cultivar)and the desirable trait transferred from the donor parent. After theinitial cross, individuals possessing the phenotype of the donor parentare selected and repeatedly crossed (backcrossed) to the recurrentparent. The resulting plant is expected to have the attributes of therecurrent parent (e.g., cultivar) and the desirable trait transferredfrom the donor parent.

The single-seed descent procedure in the strict sense refers to plantinga segregating population, harvesting a sample of one seed per plant, andusing the one-seed sample to plant the next generation. When thepopulation has been advanced from the F2 to the desired level ofinbreeding, the plants from which lines are derived will each trace todifferent F2 individuals. The number of plants in a population declineseach generation due to failure of some seeds to germinate or some plantsto produce at least one seed. As a result, not all of the F2 plantsoriginally sampled in the population will be represented by a progenywhen generation advance is completed.

In a multiple-seed procedure, breeders commonly harvest one or morefruit containing seed from each plant in a population and blend themtogether to form a bulk seed lot. Part of the bulked seed is used toplant the next generation and part is put in reserve. The procedure hasbeen referred to as modified single-seed descent or the bulk technique.

The multiple-seed procedure has been used to save labor at harvest. Itis considerably faster than removing one seed from each fruit by handfor the single seed procedure. The multiple-seed procedure also makes itpossible to plant the same number of seeds of a population eachgeneration of inbreeding. Enough seeds are harvested to make up forthose plants that did not germinate or produce seed.

Descriptions of other breeding methods that are commonly used fordifferent traits and crops can be found in one of several referencebooks (e.g., R. W. Allard, 1960, Principles of Plant Breeding, JohnWiley and Son, pp. 115-161; N. W. Simmonds, 1979, Principles of CropImprovement, Longman Group Limited; W. R. Fehr, 1987, Principles of CropDevelopment, Macmillan Publishing Co.; N. F. Jensen, 1988, PlantBreeding Methodology, John Wiley & Sons).

When the term hybrid cantaloupe plant is used in the context of thepresent invention, this also includes any hybrid cantaloupe plant whereone or more desired trait has been introduced through backcrossingmethods, whether such trait is a naturally occurring one, a mutant or atransgenic one. Backcrossing methods can be used with the presentinvention to improve or introduce one or more characteristic into theinbred parental line of the hybrid cantaloupe plant of the presentinvention. The term “backcrossing” as used herein refers to the repeatedcrossing of a hybrid progeny back to the recurrent parent, i.e.,backcrossing one, two, three, four, five, six, seven, eight, nine, ormore times to the recurrent parent. The parental cantaloupe plant whichcontributes the gene or the genes for the desired characteristic istermed the nonrecurrent or donor parent. This terminology refers to thefact that the nonrecurrent parent is used one time in the backcrossprotocol and therefore does not recur. The parental cantaloupe plant towhich the gene or genes from the nonrecurrent parent are transferred isknown as the recurrent parent as it is used for several rounds in thebackcrossing protocol.

In a typical backcross protocol, the original inbred of interest(recurrent parent) is crossed to a second inbred (nonrecurrent parent)that carries the gene or genes of interest to be transferred. Theresulting progeny from this cross are then crossed again to therecurrent parent and the process is repeated until a cantaloupe plant isobtained wherein all the desired morphological and physiologicalcharacteristics of the recurrent parent are recovered in the convertedplant, generally determined at a 5% significance level when grown in thesame environmental conditions, in addition to the gene or genestransferred from the nonrecurrent parent. It has to be noted that some,one, two, three or more, self-pollination and growing of populationmight be included between two successive backcrosses. Indeed, anappropriate selection in the population produced by theself-pollination, i.e. selection for the desired trait and physiologicaland morphological characteristics of the recurrent parent might beequivalent to one, two or even three additional backcrosses in acontinuous series without rigorous selection, saving then time, moneyand effort to the breeder. A non-limiting example of such a protocolwould be the following: a) the first generation F1 produced by the crossof the recurrent parent A by the donor parent B is backcrossed to parentA, b) selection is practiced for the plants having the desired trait ofparent B, c) selected plant are self-pollinated to produce a populationof plants where selection is practiced for the plants having the desiredtrait of parent B and physiological and morphological characteristics ofparent A, d) the selected plants are backcrossed one, two, three, four,five, six, seven, eight, nine, or more times to parent A to produceselected backcross progeny plants comprising the desired trait of parentB and the physiological and morphological characteristics of parent A.Step (c) may or may not be repeated and included between the backcrossesof step (d).

The selection of a suitable recurrent parent is an important step for asuccessful backcrossing procedure. The goal of a backcross protocol isto alter or substitute one or more trait(s) or characteristic(s) in theoriginal inbred parental line in order to find it then in the hybridmade thereof. To accomplish this, a gene or genes of the recurrentinbred is modified or substituted with the desired gene or genes fromthe nonrecurrent parent, while retaining essentially all of the rest ofthe desired genetic, and therefore the desired physiological andmorphological, constitution of the original inbred. The choice of theparticular nonrecurrent parent will depend on the purpose of thebackcross; one of the major purposes is to add some commerciallydesirable, agronomically important trait(s) to the plant. The exactbackcrossing protocol will depend on the characteristic(s) or trait(s)being altered to determine an appropriate testing protocol. Althoughbackcrossing methods are simplified when the characteristic beingtransferred is a single gene and dominant allele, multiple genes andrecessive allele(s) may also be transferred and therefore, backcrossbreeding is by no means restricted to character(s) governed by one or afew genes. In fact the number of genes might be less important that theidentification of the character(s) in the segregating population. Inthis instance it may then be necessary to introduce a test of theprogeny to determine if the desired characteristic(s) has beensuccessfully transferred. Such tests encompass visual inspection, simplecrossing, but also follow up of the characteristic(s) throughgenetically associated markers and molecular assisted breeding tools.For example, selection of progeny containing the transferred trait isdone by direct selection, visual inspection for a trait associated witha dominant allele, while the selection of progeny for a trait that istransferred via a recessive allele, such as the waxy starchcharacteristic in corn, require selfing the progeny to determine whichplant carry the recessive allele(s).

Many single gene traits have been identified that are not regularlyselected for in the development of a new parental inbred of a hybridcantaloupe plant according to the invention but that can be improved bybackcrossing techniques. Single gene traits may or may not betransgenic. Examples of these traits include but are not limited to,male sterility (such as a PR glucanase gene or the ms1, ms2, ms3, ms4 orms5 genes), herbicide resistance (such as bar or PAT genes), gynoecia(such as the g gene), resistance for bacterial, fungal (genes Fom-1 andFom-2 for resistance to fusarium wilt), or viral disease (gene nsv forresistance to melon necrotic spot virus, gene ZYM for the resistance tothe zucchini yellow mosaic virus), insect resistance (gene Vat forresistance to Aphis gossypii), male fertility, enhanced nutritionalquality, enhanced sugar content, yield stability and yield enhancement.These genes are generally inherited through the nucleus. Some knownexceptions to this are the genes for male sterility, some of which areinherited cytoplasmically, but still act as single gene traits. Severalof these single gene traits are described in U.S. Pat. Nos. 5,777,196;5,948,957 and 5,969,212, the disclosures of which are specificallyhereby incorporated by reference.

In 1981, the backcross method of breeding counted for 17% of the totalbreeding effort for inbred line development in the United States,accordingly to, Hallauer, A. R. et al. (1988) “Corn Breeding” Corn andCorn Improvement, No. 18, pp. 463-481.

The backcross breeding method provides a precise way of improvingvarieties that excel in a large number of attributes but are deficientin a few characteristics. (Page 150 of the Pr. R. W. Allard's 1960 book,published by John Wiley & Sons, Inc, Principles of Plant Breeding). Themethod makes use of a series of backcrosses to the variety to beimproved during which the character or the characters in whichimprovement is sought is maintained by selection. At the end of thebackcrossing the gene or genes being transferred unlike all other genes,will be heterozygous. Selfing after the last backcross produceshomozygosity for this gene pair(s) and, coupled with selection, willresult in a parental line of a hybrid variety with exactly theadaptation, yielding ability and quality characteristics of therecurrent parent but superior to that parent in the particularcharacteristic(s) for which the improvement program was undertaken.Therefore, this method provides the plant breeder with a high degree ofgenetic control of his work.

The method is scientifically exact because the morphological andagricultural features of the improved variety could be described inadvance and because the same variety could, if it were desired, be breda second time by retracing the same steps (Briggs, “Breeding wheatsresistant to bunt by the backcross method”, 1930 Jour. Amer. Soc.Agron., 22: 289-244).

Backcrossing is a powerful mechanism for achieving homozygosity and anypopulation obtained by backcrossing must rapidly converge on thegenotype of the recurrent parent. When backcrossing is made the basis ofa plant breeding program, the genotype of the recurrent parent will bemodified only with regards to genes being transferred, which aremaintained in the population by selection.

Successful backcrosses are, for example, the transfer of stem rustresistance from ‘Hope’ wheat to ‘Bart wheat’ and even pursuing thebackcrosses with the transfer of bunt resistance to create ‘Bart 38’,having both resistances. Also highlighted by Allard is the successfultransfer of mildew, leaf spot and wilt resistances in California Commonalfalfa to create ‘Caliverde’. This new ‘Caliverde’ variety producedthrough the backcross process is indistinguishable from CaliforniaCommon except for its resistance to the three named diseases.

One of the advantages of the backcross method is that the breedingprogram can be carried out in almost every environment that will allowthe development of the character being transferred.

The backcross technique is not only desirable when breeding for diseaseresistance but also for the adjustment of morphological characters,colour characteristics and simply inherited quantitative characters suchas earliness, plant height and seed size and shape. In this regard, amedium grain type variety, ‘Calady’, has been produced by Jones andDavis. As dealing with quantitative characteristics, they selected thedonor parent with the view of sacrificing some of the intensity of thecharacter for which it was chosen, i.e. grain size. ‘Lady Wright’, along grain variety was used as the donor parent and ‘Coloro’, a shortgrain one as the recurrent parent. After four backcrosses, the mediumgrain type variety ‘Calady’ was produced.

iii Open-Pollinated Populations

The improvement of open-pollinated populations of such crops as rye,many maizes and sugar beets, herbage grasses, legumes such as alfalfaand clover, and tropical tree crops such as cacao, coconuts, oil palmand some rubber, depends essentially upon changing gene-frequenciestowards fixation of favorable alleles while maintaining a high (but farfrom maximal) degree of heterozygosity.

Uniformity in such populations is impossible and trueness-to-type in anopen-pollinated variety is a statistical feature of the population as awhole, not a characteristic of individual plants. Thus, theheterogeneity of open-pollinated populations contrasts with thehomogeneity (or virtually so) of inbred lines, clones and hybrids.

Population improvement methods fall naturally into two groups, thosebased on purely phenotypic selection, normally called mass selection,and those based on selection with progeny testing. Interpopulationimprovement utilizes the concept of open breeding populations; allowinggenes to flow from one population to another. Plants in one population(cultivar, strain, ecotype, or any germplasm source) are crossed eithernaturally (e.g., by wind) or by hand or by bees (commonly Apis melliferaL. or Megachile rotundata F.) with plants from other populations.Selection is applied to improve one (or sometimes both) population(s) byisolating plants with desirable traits from both sources.

There are basically two primary methods of open-pollinated populationimprovement.

First, there is the situation in which a population is changed en masseby a chosen selection procedure. The outcome is an improved populationthat is indefinitely propagable by random-mating within itself inisolation.

Second, the synthetic variety attains the same end result as populationimprovement, but is not itself propagable as such; it has to bereconstructed from parental lines or clones. These plant breedingprocedures for improving open-pollinated populations are well known tothose skilled in the art and comprehensive reviews of breedingprocedures routinely used for improving cross-pollinated plants areprovided in numerous texts and articles, including: Allard, Principlesof Plant Breeding, John Wiley & Sons, Inc. (1960); Simmonds, Principlesof Crop Improvement, Longman Group Limited (1979); Hallauer and Miranda,Quantitative Genetics in Maize Breeding, Iowa State University Press(1981); and, Jensen, Plant Breeding Methodology, John Wiley & Sons, Inc.(1988).

A) Mass Selection

Mass and recurrent selections can be used to improve populations ofeither self- or cross-pollinating crops. A genetically variablepopulation of heterozygous individuals is either identified or createdby intercrossing several different parents. The best plants are selectedbased on individual superiority, outstanding progeny, or excellentcombining ability. The selected plants are intercrossed to produce a newpopulation in which further cycles of selection are continued. In massselection, desirable individual plants are chosen, harvested, and theseed composited without progeny testing to produce the followinggeneration. Since selection is based on the maternal parent only, andthere is no control over pollination, mass selection amounts to a formof random mating with selection. As stated above, the purpose of massselection is to increase the proportion of superior genotypes in thepopulation.

B) Synthetics

A synthetic variety is produced by crossing inter se a number ofgenotypes selected for good combining ability in all possible hybridcombinations, with subsequent maintenance of the variety by openpollination. Whether parents are (more or less inbred) seed-propagatedlines, as in some sugar beet and beans (Vicia) or clones, as in herbagegrasses, clovers and alfalfa, makes no difference in principle. Parentsare selected on general combining ability, sometimes by test crosses ortoperosses, more generally by polycrosses. Parental seed lines may bedeliberately inbred (e.g. by selfing or sib crossing). However, even ifthe parents are not deliberately inbred, selection within lines duringline maintenance will ensure that some inbreeding occurs. Clonal parentswill, of course, remain unchanged and highly heterozygous.

Whether a synthetic can go straight from the parental seed productionplot to the farmer or must first undergo one or more cycles ofmultiplication depends on seed production and the scale of demand forseed. In practice, grasses and clovers are generally multiplied once ortwice and are thus considerably removed from the original synthetic.

While mass selection is sometimes used, progeny testing is generallypreferred for polycrosses, because of their operational simplicity andobvious relevance to the objective, namely exploitation of generalcombining ability in a synthetic.

The number of parental lines or clones that enters a synthetic varieswidely. In practice, numbers of parental lines range from 10 to severalhundred, with 100-200 being the average. Broad based synthetics formedfrom 100 or more clones would be expected to be more stable during seedmultiplication than narrow based synthetics.

iv. Hybrids

A hybrid is an individual plant resulting from a cross between parentsof differing genotypes. Commercial hybrids are now used extensively inmany crops, including corn (maize), sorghum, sugarbeet, sunflower andbroccoli. Hybrids can be formed in a number of different ways, includingby crossing two parents directly (single cross hybrids), by crossing asingle cross hybrid with another parent (three-way or triple crosshybrids), or by crossing two different hybrids (four-way or double crosshybrids).

Strictly speaking, most individuals in an out breeding (i.e.,open-pollinated) population are hybrids, but the term is usuallyreserved for cases in which the parents are individuals whose genomesare sufficiently distinct for them to be recognized as different speciesor subspecies. Hybrids may be fertile or sterile depending onqualitative and/or quantitative differences in the genomes of the twoparents. Heterosis, or hybrid vigor, is usually associated withincreased heterozygosity that results in increased vigor of growth,survival, and fertility of hybrids as compared with the parental linesthat were used to form the hybrid. Maximum heterosis is usually achievedby crossing two genetically different, highly inbred lines.

Hybrid commercial cantaloupe seed is produced by hand pollination.Pollen of the male parent is harvested and manually applied to thestigmatic surface of the female inbred. Prior to, and after handpollination, flowers are covered so that insects do not bring foreignpollen and create a mix or impurity. Flowers are tagged to identifypollinated fruit from which seed will be harvested.

Once the inbreds that give the best hybrid performance have beenidentified, the hybrid seed can be reproduced indefinitely as long asthe homogeneity of the inbred parent is maintained. A single-crosshybrid is produced when two inbred lines are crossed to produce the F1progeny. A double-cross hybrid is produced from four inbred linescrossed in pairs (A×B and C×D) and then the two F1 hybrids are crossedagain (A×B)×(C×D). Much of the hybrid vigor and uniformity exhibited byF1 hybrids is lost in the next generation (F2). Consequently, seed fromF2 hybrid varieties is not used for planting stock.

The production of hybrids is a well-developed industry, involving theisolated production of both the parental lines and the hybrids whichresult from crossing those lines. For a detailed discussion of thehybrid production process, see, e.g., Wright, Commercial Hybrid SeedProduction 8:161-176, In Hybridization of Crop Plants.

v. Bulk Segregation Analysis (BSA)

BSA, a.k.a. bulked segregation analysis, or bulk segregant analysis, isa method described by Michelmore et al. (Michelmore et al., 1991,Identification of markers linked to disease-resistance genes by bulkedsegregant analysis: a rapid method to detect markers in specific genomicregions by using segregating populations. Proceedings of the NationalAcademy of Sciences, USA, 99:9828-9832) and Quarrie et al. (Quarrie etal., 1999, Journal of Experimental Botany, 50(337):1299-1306).

For BSA of a trait of interest, parental lines with certain differentphenotypes are chosen and crossed to generate F2, doubled haploid orrecombinant inbred populations with QTL analysis. The population is thenphenotyped to identify individual plants or lines having high or lowexpression of the trait. Two DNA bulks are prepared, one from theindividuals having one phenotype (e.g., resistant to virus), and theother from the individuals having reversed phenotype (e.g., susceptibleto virus), and analyzed for allele frequency with molecular markers.Only a few individuals are required in each bulk (e.g., 10 plants each)if the markers are dominant (e.g., RAPDs). More individuals are neededwhen markers are co-dominant (e.g., RFLPs). Markers linked to thephenotype can be identified and used for breeding or QTL mapping.

vi. Hand-Pollination Method

Hand pollination describes the crossing of plants via the deliberatefertilization of female ovules with pollen from a desired male parentplant. In some embodiments the donor or recipient female parent and thedonor or recipient male parent line are planted in the same field. Theinbred male parent can be planted earlier than the female parent toensure adequate pollen supply at the pollination time. In someembodiments, the male parent and female parent can be planted at a ratioof 1 male parent to 4-10 female parents. The diploid male parent may beplanted at the top of the field for efficient male flower collectionduring pollination. Pollination is started when the female parent floweris ready to be fertilized. Female flower buds that are ready to open inthe following days are identified, covered with paper cups or smallpaper bags that prevent bee or any other insect from visiting the femaleflowers, and marked with any kind of material that can be easily seenthe next morning. In some embodiments, this process is best done in theafternoon. The male flowers of the diploid male parent are collected inthe early morning before they are open and visited by pollinatinginsects. The covered female flowers of the female parent, which haveopened, are un-covered and pollinated with the collected fresh maleflowers of the diploid male parent, starting as soon as the male flowersheds pollen. The pollinated female flowers are again covered afterpollination to prevent bees and any other insects visit. The pollinatedfemale flowers are also marked. The marked fruits are harvested. In someembodiments, the male pollen used for fertilization has been previouslycollected and stored.

vii. Bee-Pollination Method

Using the bee-pollination method, the parent plants are usually plantedwithin close proximity. In some embodiments more female plants areplanted to allow for a greater production of seed. Breeding of dioeciousspecies can also be done by growing equal amount of each parent plant.Beehives are placed in the field for transfer of pollen by bees from themale parent to the female flowers of the female parent. In someembodiments, fruits set after the introduction of the beehives can bemarked for later collection.

viii. Targeting Induced Local Lesions in Genomes (TILLING)

Breeding schemes of the present application can include crosses withTILLING® plant lines. TILLING® is a method in molecular biology thatallows directed identification of mutations in a specific gene. TILLING®was introduced in 2000, using the model plant Arabidopsis thaliana.TILLING® has since been used as a reverse genetics method in otherorganisms such as zebrafish, corn, wheat, rice, soybean, tomato andlettuce.

The method combines a standard and efficient technique of mutagenesiswith a chemical mutagen (e.g., Ethyl methanesulfonate (EMS)) with asensitive DNA screening-technique that identifies single base mutations(also called point mutations) in a target gene. EcoTILLING is a methodthat uses TILLING® techniques to look for natural mutations inindividuals, usually for population genetics analysis (see Comai, etal., 2003 The Plant Journal 37, 778-786; Gilchrist et al. 2006 Mol.Ecol. 15, 1367-1378; Mejlhede et al. 2006 Plant Breeding 125, 461-467;Nieto et al. 2007 BMC Plant Biology 7, 34-42, each of which isincorporated by reference hereby for all purposes). DEcoTILLING is amodification of TILLING® and EcoTILLING which uses an inexpensive methodto identify fragments (Garvin et al., 2007, DEco-TILLING: An inexpensivemethod for SNP discovery that reduces ascertainment bias. MolecularEcology Notes 7, 735-746).

The TILLING® method relies on the formation of heteroduplexes that areformed when multiple alleles (which could be from a heterozygote or apool of multiple homozygotes and heterozygotes) are amplified in a PCR,heated, and then slowly cooled. A “bubble” forms at the mismatch of thetwo DNA strands (the induced mutation in TILLING® or the naturalmutation or SNP in EcoTILLING), which is then cleaved by single strandednucleases. The products are then separated by size on several differentplatforms.

Several TILLING® centers exists over the world that focus onagriculturally important species: UC Davis (USA), focusing on Rice;Purdue University (USA), focusing on Maize; University of BritishColumbia (CA), focusing on Brassica napus; John Innes Centre (UK),focusing on Brassica rapa; Fred Hutchinson Cancer Research, focusing onArabidopsis; Southern Illinois University (USA), focusing on Soybean;John Innes Centre (UK), focusing on Lotus and Medicago; and INRA(France), focusing on Pea and Tomato.

More detailed description on methods and compositions on TILLING® can befound in U.S. Pat. No. 5,994,075, US 2004/0053236 A1, WO 2005/055704,and WO 2005/048692, each of which is hereby incorporated by referencefor all purposes.

Thus in some embodiments, the breeding methods of the present disclosureinclude breeding with one or more TILLING plant lines with one or moreidentified mutations.

viii Mutation Breeding

Mutation breeding is another method of introducing new traits intocantaloupe plants. Mutations that occur spontaneously or areartificially induced can be useful sources of variability for a plantbreeder. The goal of artificial mutagenesis is to increase the rate ofmutation for a desired characteristic. Mutation rates can be increasedby many different means or mutating agents including temperature,long-term seed storage, tissue culture conditions, radiation (such asX-rays, Gamma rays, neutrons, Beta radiation, or ultraviolet radiation),chemical mutagens (such as base analogs like 5-bromo-uracil),antibiotics, alkylating agents (such as sulfur mustards, nitrogenmustards, epoxides, ethyleneamines, sulfates, sulfonates, sulfones, orlactones), azide, hydroxylamine, nitrous acid or acridines. Once adesired trait is observed through mutagenesis the trait may then beincorporated into existing germplasm by traditional breeding techniques.Details of mutation breeding can be found in W. R. Fehr, 1993,Principles of Cultivar Development, Macmillan Publishing Co.

New breeding techniques such as the ones involving the uses of ZincFinger Nucleases or oligonucleotide directed mutagenesis shall also beused to generate genetic variability and introduce new traits intocantaloupe varieties.

ix. Double Haploids and Chromosome Doubling

One way to obtain homozygous plants without the need to cross twoparental lines followed by a long selection of the segregating progeny,and/or multiple back-crossings is to produce haploids and then doublethe chromosomes to form doubled haploids. Haploid plants can occurspontaneously, or may be artificially induced via chemical treatments orby crossing plants with inducer lines (Seymour et al. 2012, PNAS vol109, pg 4227-4232; Zhang et al., 2008 Plant Cell Rep. Dec. 27(12)1851-60). The production of haploid progeny can occur via a variety ofmechanisms which can affect the distribution of chromosomes duringgamete formation. The chromosome complements of haploids sometimesdouble spontaneously to produce homozygous doubled haploids (DH5).Mixoploids, which are plants which contain cells having differentploidies, can sometimes arise and may represent plants that areundergoing chromosome doubling so as to spontaneously produce doubledhaploid tissues, organs, shoots, floral parts or plants. Another commontechnique is to induce the formation of double haploid plants with achromosome doubling treatment such as colchicine (El-Hennawy et al.,2011 Vol 56, issue 2 pg 63-72; Doubled Haploid Production in Crop Plants2003 edited by Maluszynski ISBN 1-4020-1544-5). The production ofdoubled haploid plants yields highly uniform inbred lines and isespecially desirable as an alternative to sexual inbreeding oflonger-generation crops. By producing doubled haploid progeny, thenumber of possible gene combinations for inherited traits is moremanageable. Thus, an efficient doubled haploid technology cansignificantly reduce the time and the cost of inbred and cultivardevelopment.

x. Protoplast Fusion

In another method for breeding plants, protoplast fusion can also beused for the transfer of trait-conferring genomic material from a donorplant to a recipient plant. Protoplast fusion is an induced orspontaneous union, such as a somatic hybridization, between two or moreprotoplasts (cells of which the cell walls are removed by enzymatictreatment) to produce a single bi- or multi-nucleate cell. The fusedcell that may even be obtained with plant species that cannot beinterbred in nature is tissue cultured into a hybrid plant exhibitingthe desirable combination of traits.

xi. Embryo Rescue

Alternatively, embryo rescue may be employed in the transfer ofresistance-conferring genomic material from a donor plant to a recipientplant. Embryo rescue can be used as a procedure to isolate embryo's fromcrosses wherein plants fail to produce viable seed. In this process, thefertilized ovary or immature seed of a plant is tissue cultured tocreate new plants (see Pierik, 1999, In vitro culture of higher plants,Springer, ISBN 079235267x, 9780792352679, which is incorporated hereinby reference in its entirety).

Breeding Evaluation

Each breeding program can include a periodic, objective evaluation ofthe efficiency of the breeding procedure. Evaluation criteria varydepending on the goal and objectives, but should include gain fromselection per year based on comparisons to an appropriate standard,overall value of the advanced breeding lines, and number of successfulcultivars produced per unit of input (e.g., per year, per dollarexpended, etc.).

Promising advanced breeding lines are thoroughly tested per se and inhybrid combination and compared to appropriate standards in environmentsrepresentative of the commercial target area(s). The best lines arecandidates for use as parents in new commercial cultivars; those stilldeficient in a few traits may be used as parents to produce newpopulations for further selection.

In one embodiment, the plants are selected on the basis of one or morephenotypic traits. Skilled persons will readily appreciate that suchtraits include any observable characteristic of the plant, including forexample growth rate, height, weight, color, taste, smell, changes in theproduction of one or more compounds by the plant (including for example,metabolites, proteins, drugs, carbohydrates, oils, and any othercompounds).

A most difficult task is the identification of individuals that aregenetically superior, because for most traits the true genotypic valueis masked by other confounding plant traits or environmental factors.One method of identifying a superior plant is to observe its performancerelative to other experimental plants and to a widely grown standardcultivar. If a single observation is inconclusive, replicatedobservations provide a better estimate of its genetic worth.

Proper testing should detect any major faults and establish the level ofsuperiority or improvement over current cultivars. In addition toshowing superior performance, there must be a demand for a new cultivarthat is compatible with industry standards or which creates a newmarket. The introduction of a new cultivar will incur additional coststo the seed producer, the grower, processor and consumer; for specialadvertising and marketing, altered seed and commercial productionpractices, and new product utilization. The testing preceding release ofa new cultivar should take into consideration research and developmentcosts as well as technical superiority of the final cultivar. Forseed-propagated cultivars, it must be feasible to produce seed easilyand economically.

It should be appreciated that in certain embodiments, plants may beselected based on the absence, suppression or inhibition of a certainfeature or trait (such as an undesirable feature or trait) as opposed tothe presence of a certain feature or trait (such as a desirable featureor trait).

Selecting plants based on genotypic information is also envisaged (forexample, including the pattern of plant gene expression, genotype, orpresence of genetic markers). Where the presence of one or more geneticmarker is assessed, the one or more marker may already be known and/orassociated with a particular characteristic of a plant; for example, amarker or markers may be associated with an increased growth rate ormetabolite profile. This information could be used in combination withassessment based on other characteristics in a method of the disclosureto select for a combination of different plant characteristics that maybe desirable. Such techniques may be used to identify novel quantitativetrait loci (QTLs). By way of example, plants may be selected based ongrowth rate, size (including but not limited to weight, height, leafsize, stem size, branching pattern, or the size of any part of theplant), general health, survival, tolerance to adverse physicalenvironments and/or any other characteristic, as described hereinbefore.

Further non-limiting examples include selecting plants based on: speedof seed germination; quantity of biomass produced; increased root,and/or leaf/shoot growth that leads to an increased yield (herbage orgrain or fiber or oil) or biomass production; effects on plant growththat results in an increased seed yield for a crop; effects on plantgrowth which result in an increased fruit yield; effects on plant growththat lead to an increased resistance or tolerance disease includingfungal, viral or bacterial diseases or to pests such as insects, mitesor nematodes in which damage is measured by decreased foliar symptomssuch as the incidence of bacterial or fungal lesions, or area of damagedfoliage or reduction in the numbers of nematode cysts or galls on plantroots, or improvements in plant yield in the presence of such plantpests and diseases; effects on plant growth that lead to increasedmetabolite yields; effects on plant growth that lead to improvedaesthetic appeal which may be particularly important in plants grown fortheir form, color or taste, for example the color intensity and form ofcantaloupe rinds or flesh, or the taste of said fruit.

Molecular Breeding Evaluation Techniques

Selection of plants based on phenotypic or genotypic information may beperformed using techniques such as, but not limited to: high through-putscreening of chemical components of plant origin, sequencing techniquesincluding high through-put sequencing of genetic material, differentialdisplay techniques (including DDRT-PCR, and DD-PCR), nucleic acidmicroarray techniques, RNA-seq (Whole Transcriptome Shotgun Sequencing),qRTPCR (quantitative real time PCR).

In one embodiment, the evaluating step of a plant breeding programinvolves the identification of desirable traits in progeny plants.Progeny plants can be grown in, or exposed to conditions designed toemphasize a particular trait (e.g. drought conditions for droughttolerance, lower temperatures for freezing tolerant traits). Progenyplants with the highest scores for a particular trait may be used forsubsequent breeding steps.

In some embodiments, plants selected from the evaluation step canexhibit a 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%,65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 120% or more improvement in aparticular plant trait compared to a control plant.

In other embodiments, the evaluating step of plant breeding comprisesone or more molecular biological tests for genes or other markers. Forexample, the molecular biological test can involve probe hybridizationand/or amplification of nucleic acid (e.g., measuring nucleic aciddensity by Northern or Southern hybridization, PCR) and/or immunologicaldetection (e.g., measuring protein density, such as precipitation andagglutination tests, ELISA (e.g., Lateral Flow test or DAS-ELISA),Western blot, RIA, immune labeling, immunosorbent electron microscopy(ISEM), and/or dot blot).

The procedure to perform a nucleic acid hybridization, an amplificationof nucleic acid (e.g., RT-PCR) or an immunological detection (e.g.,precipitation and agglutination tests, ELISA (e.g., Lateral Flow test orDAS-ELISA), Western blot, RIA, immunogold or immunofluorescent labeling,immunosorbent electron microscopy (ISEM), and/or dot blot tests) areperformed as described elsewhere herein and well-known by one skilled inthe art.

In one embodiment, the evaluating step comprises PCR (semi-quantitativeor quantitative), wherein primers are used to amplify one or morenucleic acid sequences of a desirable gene, or a nucleic acid associatedwith said gene or a desirable trait (e.g., a co-segregating nucleicacid, or other marker).

In another embodiment, the evaluating step comprises immunologicaldetection (e.g., precipitation and agglutination tests, ELISA (e.g.,Lateral Flow test or DAS-ELISA), Western blot, RIA, immuno labeling(gold, fluorescent, or other detectable marker), immunosorbent electronmicroscopy (ISEM), and/or dot blot), wherein one or more gene ormarker-specific antibodies are used to detect one or more desirableproteins. In one embodiment, said specific antibody is selected from thegroup consisting of polyclonal antibodies, monoclonal antibodies,antibody fragments, and combination thereof.

Reverse Transcription Polymerase Chain Reaction (RT-PCR) can be utilizedin the present disclosure to determine expression of a gene to assistduring the selection step of a breeding scheme. It is a variant ofpolymerase chain reaction (PCR), a laboratory technique commonly used inmolecular biology to generate many copies of a DNA sequence, a processtermed “amplification”. In RT-PCR, however, RNA strand is first reversetranscribed into its DNA complement (complementary DNA, or cDNA) usingthe enzyme reverse transcriptase, and the resulting cDNA is amplifiedusing traditional or real-time PCR.

RT-PCR utilizes a pair of primers, which are complementary to a definedsequence on each of the two strands of the cDNA. These primers are thenextended by a DNA polymerase and a copy of the strand is made after eachcycle, leading to logarithmic amplification.

RT-PCR includes three major steps. The first step is the reversetranscription (RT) where RNA is reverse transcribed to cDNA using areverse transcriptase and primers. This step is very important in orderto allow the performance of PCR since DNA polymerase can act only on DNAtemplates. The RT step can be performed either in the same tube with PCR(one-step PCR) or in a separate one (two-step PCR) using a temperaturebetween 40° C. and 50° C., depending on the properties of the reversetranscriptase used.

The next step involves the denaturation of the dsDNA at 95° C., so thatthe two strands separate and the primers can bind again at lowertemperatures and begin a new chain reaction. Then, the temperature isdecreased until it reaches the annealing temperature which can varydepending on the set of primers used, their concentration, the probe andits concentration (if used), and the cations concentration. The mainconsideration, of course, when choosing the optimal annealingtemperature is the melting temperature (Tm) of the primers and probes(if used). The annealing temperature chosen for a PCR depends directlyon length and composition of the primers. This is the result of thedifference of hydrogen bonds between A-T (2 bonds) and G-C (3 bonds). Anannealing temperature about 5 degrees below the lowest Tm of the pair ofprimers is usually used.

The final step of PCR amplification is the DNA extension from theprimers which is done by the thermostable Taq DNA polymerase usually at72° C., which is the optimal temperature for the polymerase to work. Thelength of the incubation at each temperature, the temperaturealterations and the number of cycles are controlled by a programmablethermal cycler. The analysis of the PCR products depends on the type ofPCR applied. If a conventional PCR is used, the PCR product is detectedusing agarose gel electrophoresis and ethidium bromide (or other nucleicacid staining)

Conventional RT-PCR is a time-consuming technique with importantlimitations when compared to real time PCR techniques. This, combinedwith the fact that ethidium bromide has low sensitivity, yields resultsthat are not always reliable. Moreover, there is an increasedcross-contamination risk of the samples since detection of the PCRproduct requires the post-amplification processing of the samples.Furthermore, the specificity of the assay is mainly determined by theprimers, which can give false-positive results. However, the mostimportant issue concerning conventional RT-PCR is the fact that it is asemi or even a low quantitative technique, where the amplicon can bevisualized only after the amplification ends.

Real time RT-PCR provides a method where the amplicons can be visualizedas the amplification progresses using a fluorescent reporter molecule.There are three major kinds of fluorescent reporters used in real timeRT-PCR, general non specific DNA Binding Dyes such as SYBR Green I,TaqMan Probes and Molecular Beacons (including Scorpions).

The real time PCR thermal cycler has a fluorescence detection threshold,below which it cannot discriminate the difference between amplificationgenerated signal and background noise. On the other hand, thefluorescence increases as the amplification progresses and theinstrument performs data acquisition during the annealing step of eachcycle. The number of amplicons will reach the detection baseline after aspecific cycle, which depends on the initial concentration of the targetDNA sequence. The cycle at which the instrument can discriminate theamplification generated fluorescence from the background noise is calledthe threshold cycle (Ct). The higher is the initial DNA concentration,the lower its Ct will be.

Other forms of nucleic acid detection can include next generationsequencing methods such as DNA SEQ or RNA SEQ using any known sequencingplatform including, but not limited to: Roche 454, Solexa GenomeAnalyzer, AB SOLiD, Illumina GA/HiSeq, Ion PGM, Mi Seq, among others(Liu et al., 2012 Journal of Biomedicine and Biotechnology Volume 2012ID 251364; Franca et al., 2002 Quarterly Reviews of Biophysics 35 pg169-200; Mardis 2008 Genomics and Human Genetics vol 9 pg 387-402).

In other embodiments, nucleic acids may be detected with other highthroughput hybridization technologies including microarrays, gene chips,LNA probes, nanoStrings, and fluorescence polarization detection amongothers.

In some embodiments, detection of markers can be achieved at an earlystage of plant growth by harvesting a small tissue sample (e.g., branch,or leaf disk). This approach is preferable when working with largepopulations as it allows breeders to weed out undesirable progeny at anearly stage and conserve growth space and resources for progeny whichshow more promise. In some embodiments the detection of markers isautomated, such that the detection and storage of marker data is handledby a machine. Recent advances in robotics have also led to full serviceanalysis tools capable of handling nucleic acid/protein markerextractions, detection, storage and analysis.

Quantitative Trait Loci

Breeding schemes of the present application can include crosses betweendonor and recipient plants. In some embodiments said donor plantscontain a gene or genes of interest which may confer the plant with adesirable phenotype. The recipient line can be an elite line havingcertain favorite traits such for commercial production. In oneembodiment, the elite line may contain other genes that also impart saidline with the desired phenotype. When crossed together, the donor andrecipient plant may create a progeny plant with combined desirable lociwhich may provide quantitatively additive effect of a particularcharacteristic. In that case, QTL mapping can be involved to facilitatethe breeding process.

A QTL (quantitative trait locus) mapping can be applied to determine theparts of the donor plant's genome conferring the desirable phenotype,and facilitate the breeding methods. Inheritance of quantitative traitsor polygenic inheritance refers to the inheritance of a phenotypiccharacteristic that varies in degree and can be attributed to theinteractions between two or more genes and their environment. Though notnecessarily genes themselves, quantitative trait loci (QTLs) arestretches of DNA that are closely linked to the genes that underlie thetrait in question. QTLs can be molecularly identified to help mapregions of the genome that contain genes involved in specifying aquantitative trait. This can be an early step in identifying andsequencing these genes.

Typically, QTLs underlie continuous traits (those traits that varycontinuously, e.g. yield, height, level of resistance to virus, etc.) asopposed to discrete traits (traits that have two or several charactervalues, e.g. smooth vs. wrinkled peas used by Mendel in hisexperiments). Moreover, a single phenotypic trait is usually determinedby many genes. Consequently, many QTLs are associated with a singletrait.

A quantitative trait locus (QTL) is a region of DNA that is associatedwith a particular phenotypic trait—these QTLs are often found ondifferent chromosomes. Knowing the number of QTLs that explainsvariation in the phenotypic trait tells about the genetic architectureof a trait. It may tell that a trait is controlled by many genes ofsmall effect, or by a few genes of large effect.

Another use of QTLs is to identify candidate genes underlying a trait.Once a region of DNA is identified as contributing to a phenotype, itcan be sequenced. The DNA sequence of any genes in this region can thenbe compared to a database of DNA for genes whose function is alreadyknown.

In a recent development, classical QTL analyses are combined with geneexpression profiling i.e. by DNA microarrays. Such expression QTLs(e-QTLs) describes cis- and trans-controlling elements for theexpression of often disease-associated genes. Observed epistatic effectshave been found beneficial to identify the gene responsible by across-validation of genes within the interacting loci with metabolicpathway- and scientific literature databases.

QTL mapping is the statistical study of the alleles that occur in alocus and the phenotypes (physical forms or traits) that they produce(see, Meksem and Kahl, The handbook of plant genome mapping: genetic andphysical mapping, 2005, Wiley-VCH, ISBN 3527311165, 9783527311163).Because most traits of interest are governed by more than one gene,defining and studying the entire locus of genes related to a trait giveshope of understanding what effect the genotype of an individual mighthave in the real world.

Statistical analysis is required to demonstrate that different genesinteract with one another and to determine whether they produce asignificant effect on the phenotype. QTLs identify a particular regionof the genome as containing a gene that is associated with the traitbeing assayed or measured. They are shown as intervals across achromosome, where the probability of association is plotted for eachmarker used in the mapping experiment.

To begin, a set of genetic markers must be developed for the species inquestion. A marker is an identifiable region of variable DNA. Biologistsare interested in understanding the genetic basis of phenotypes(physical traits). The aim is to find a marker that is significantlymore likely to co-occur with the trait than expected by chance, that is,a marker that has a statistical association with the trait. Ideally,they would be able to find the specific gene or genes in question, butthis is a long and difficult undertaking Instead, they can more readilyfind regions of DNA that are very close to the genes in question. When aQTL is found, it is often not the actual gene underlying the phenotypictrait, but rather a region of DNA that is closely linked with the gene.

For organisms whose genomes are known, one might now try to excludegenes in the identified region whose function is known with somecertainty not to be connected with the trait in question. If the genomeis not available, it may be an option to sequence the identified regionand determine the putative functions of genes by their similarity togenes with known function, usually in other genomes. This can be doneusing BLAST, an online tool that allows users to enter a primarysequence and search for similar sequences within the BLAST database ofgenes from various organisms.

Another interest of statistical geneticists using QTL mapping is todetermine the complexity of the genetic architecture underlying aphenotypic trait. For example, they may be interested in knowing whethera phenotype is shaped by many independent loci, or by a few loci, and dothose loci interact. This can provide information on how the phenotypemay be evolving.

Molecular markers are used for the visualization of differences innucleic acid sequences. This visualization is possible due to DNA-DNAhybridization techniques (RFLP) and/or due to techniques using thepolymerase chain reaction (e.g. STS, microsatellites, AFLP). Alldifferences between two parental genotypes will segregate in a mappingpopulation based on the cross of these parental genotypes. Thesegregation of the different markers may be compared and recombinationfrequencies can be calculated. The recombination frequencies ofmolecular markers on different chromosomes are generally 50%. Betweenmolecular markers located on the same chromosome the recombinationfrequency depends on the distance between the markers. A lowrecombination frequency corresponds to a low distance between markers ona chromosome. Comparing all recombination frequencies will result in themost logical order of the molecular markers on the chromosomes. Thismost logical order can be depicted in a linkage map (Paterson, 1996,Genome Mapping in Plants. R. G. Landes, Austin.). A group of adjacent orcontiguous markers on the linkage map that is associated to a reduceddisease incidence and/or a reduced lesion growth rate pinpoints theposition of a QTL.

The nucleic acid sequence of a QTL may be determined by methods known tothe skilled person. For instance, a nucleic acid sequence comprisingsaid QTL or a resistance-conferring part thereof may be isolated from adonor plant by fragmenting the genome of said plant and selecting thosefragments harboring one or more markers indicative of said QTL.Subsequently, or alternatively, the marker sequences (or parts thereof)indicative of said QTL may be used as (PCR) amplification primers, inorder to amplify a nucleic acid sequence comprising said QTL from agenomic nucleic acid sample or a genome fragment obtained from saidplant. The amplified sequence may then be purified in order to obtainthe isolated QTL. The nucleotide sequence of the QTL, and/or of anyadditional markers comprised therein, may then be obtained by standardsequencing methods.

One or more such QTLs associated with a desirable trait in a donor plantcan be transferred to a recipient plant to make incorporate thedesirable train into progeny plants by transferring and/or breedingmethods.

In one embodiment, an advanced backcross QTL analysis (AB-QTL) is usedto discover the nucleotide sequence or the QTLs responsible for theresistance of a plant. Such method was proposed by Tanksley and Nelsonin 1996 (Tanksley and Nelson, 1996, Advanced backcross QTL analysis: amethod for simultaneous discovery and transfer of valuable QTL fromun-adapted germplasm into elite breeding lines. Theor Appl Genet92:191-203) as a new breeding method that integrates the process of QTLdiscovery with variety development, by simultaneously identifying andtransferring useful QTL alleles from un-adapted (e.g., land races, wildspecies) to elite germplasm, thus broadening the genetic diversityavailable for breeding. A non-limiting exemplary scheme of AB-QTLmapping strategy is shown in FIG. 2. AB-QTL strategy was initiallydeveloped and tested in tomato, and has been adapted for use in othercrops including rice, maize, wheat, pepper, barley, and bean. Oncefavorable QTL alleles are detected, only a few additionalmarker-assisted generations are required to generate near isogenic lines(NILs) or introgression lines (ILs) that can be field tested in order toconfirm the QTL effect and subsequently used for variety development.

Isogenic lines in which favorable QTL alleles have been fixed can begenerated by systematic backcrossing and introgressing of marker-defineddonor segments in the recurrent parent background. These isogenic linesare referred as near isogenic lines (NILs), introgression lines (ILs),backcross inbred lines (BILs), backcross recombinant inbred lines(BCRIL), recombinant chromosome substitution lines (RCSLs), chromosomesegment substitution lines (CSSLs), and stepped aligned inbredrecombinant strains (STAIRSs). An introgression line in plant molecularbiology is a line of a crop species that contains genetic materialderived from a similar species. ILs represent NILs with relatively largeaverage introgression length, while BILs and BCRILs are backcrosspopulations generally containing multiple donor introgressions per line.As used herein, the term “introgression lines or ILs” refers to plantlines containing a single marker defined homozygous donor segment, andthe term “pre-ILs” refers to lines which still contain multiplehomozygous and/or heterozygous donor segments.

To enhance the rate of progress of introgression breeding, a geneticinfrastructure of exotic libraries can be developed. Such an exoticlibrary comprises of a set of introgression lines, each of which has asingle, possibly homozygous, marker-defined chromosomal segment thatoriginates from a donor exotic parent, in an otherwise homogenous elitegenetic background, so that the entire donor genome would be representedin a set of introgression lines. A collection of such introgressionlines is referred as libraries of introgression lines or IL libraries(ILLs). The lines of an ILL cover usually the complete genome of thedonor, or the part of interest. Introgression lines allow the study ofquantitative trait loci, but also the creation of new varieties byintroducing exotic traits. High resolution mapping of QTL using ILLsenable breeders to assess whether the effect on the phenotype is due toa single QTL or to several tightly linked QTL affecting the same trait.In addition, sub-ILs can be developed to discover molecular markerswhich are more tightly linked to the QTL of interest, which can be usedfor marker-assisted breeding (MAB). Multiple introgression lines can bedeveloped when the introgression of a single QTL is not sufficient toresult in a substantial improvement in agriculturally important traits(Gur and Zamir, Unused natural variation can lift yield barriers inplant breeding, 2004, PLoS Biol.; 2(10):e245).

Plant Transformation

In some embodiments, the present invention provides transformedcantaloupe plants or parts thereof that have been transformed so thatits genetic material contains one or more transgenes, preferablyoperably linked to one or more regulatory elements. Also, the inventionprovides methods for producing a cantaloupe plant containing in itsgenetic material one or more transgenes, preferably operably linked toone or more regulatory elements, by crossing transformed cantaloupeplants with a second plant of another cantaloupe, so that the geneticmaterial of the progeny that results from the cross contains thetransgene(s), preferably operably linked to one or more regulatoryelements. The invention also provides methods for producing a cantaloupeplant that contains in its genetic material one or more transgene(s),wherein the method comprises crossing a cantaloupe with a second plantof another cantaloupe which contains one or more transgene(s) operablylinked to one or more regulatory element(s) so that the genetic materialof the progeny that results from the cross contains the transgene(s)operably linked to one or more regulatory element(s). Transgeniccantaloupe plants, or parts thereof produced by the method are in thescope of the present invention.

With the advent of molecular biological techniques that have allowed theisolation and characterization of genes that encode specific proteinproducts, scientists in the field of plant biology developed a stronginterest in engineering the genome of plants to contain and expressforeign genes, or additional, or modified versions of native, orendogenous, genes (perhaps driven by different promoters) in order toalter the traits of a plant in a specific manner. Such foreignadditional and/or modified genes are referred to herein collectively as“transgenes.” Over the last fifteen to twenty years several methods forproducing transgenic plants have been developed, and the presentinvention, in particular embodiments, also relates to transformedversions of the claimed hybrid cantaloupe plant BRONCO.

Numerous methods for plant transformation have been developed, includingbiological and physical, plant transformation protocols. See, forexample, Miki et al., “Procedures for Introducing Foreign DNA intoPlants” in Methods in Plant Molecular Biology and Biotechnology, GlickB. R. and Thompson, J. E. Eds. (CRC Press, Inc., Boca Raton, 1993) pages67-88. In addition, expression vectors and in vitro culture methods forplant cell or tissue transformation and regeneration of plants areavailable. See, for example, Gruber et al., “Vectors for PlantTransformation” in Methods in Plant Molecular Biology and Biotechnology,Glick B. R. and Thompson, J. E. Eds. (CRC Press, Inc., Boca Raton, 1993)pages 89-119.

i Agrobacterium-Mediated Transformation

One method for introducing an expression vector into plants is based onthe natural transformation system of Agrobacterium. See, for example,Horsch et al., Science 227:1229 (1985), Jefferson et al., Embo J.3901-390764, (1987), Valles et al., PI. Cell. Rep. 145-148:13 (1984). A.tumefaciens and A. rhizogenes are plant pathogenic soil bacteria whichgenetically transform plant cells. The Ti and Ri plasmids of A.tumefaciens and A. rhizogenes, respectively, carry genes responsible forgenetic transformation of the plant. See, for example, Kado, C. I.,Crit. Rev. Plant Sci. 10:1 (1991). Descriptions of Agrobacterium vectorsystems and methods for Agrobacterium-mediated gene transfer areprovided by Gruber et al., supra, Miki et al., supra, and Moloney etal., Plant Cell Reports 8:238 (1989). See also, U.S. Pat. No. 5,591,616issued Jan. 7, 1997.

ii. Direct Gene Transfer

Despite the fact the host range for Agrobacterium-mediatedtransformation is broad, some major cereal crop species and gymnospermshave generally been recalcitrant to this mode of gene transfer, eventhough some success has been achieved in rice and corn. Hiei et al., ThePlant Journal 6:271-282 (1994) and U.S. Pat. No. 5,591,616 issued Jan.7, 1997. Several methods of plant transformation, collectively referredto as direct gene transfer, have been developed as an alternative toAgrobacterium-mediated transformation.

A generally applicable method of plant transformation ismicroprojectile-mediated transformation wherein DNA is carried on thesurface of microprojectiles measuring 1 to 4 micron. The expressionvector is introduced into plant tissues with a biolistic device thataccelerates the microprojectiles to speeds of 300 to 600 m/s which issufficient to penetrate plant cell walls and membranes. Sanford et al.,Part. Sci. Technol. 5:27 (1987), Sanford, J. C., Trends Biotech. 6:299(1988), Klein et al., BioTechnology 6:559-563 (1988), Sanford, J. C.,Physiol Plant 7:206 (1990), Klein et al., BioTechnology 10:268 (1992).Gray et al., Plant Cell Tissue and Organ Culture. 1994, 37:2, 179-184.

Another method for physical delivery of DNA to plants is sonication oftarget cells. Zhang et al., BioTechnology 9:996 (1991). Alternatively,liposome and spheroplast fusion have been used to introduce expressionvectors into plants. Deshayes et al., EMBO J., 4:2731 (1985), Christouet al., Proc Natl. Acad. Sci. U.S.A. 84:3962 (1987). Direct uptake ofDNA into protoplasts using CaCl₂ precipitation, polyvinyl alcohol orpoly-L-ornithine has also been reported. Hain et al., Mol. Gen. Genet.199:161 (1985) and Draper et al., Plant Cell Physiol. 23:451 (1982).Electroporation of protoplasts and whole cells and tissues have alsobeen described. D'Halluin et al., Plant Cell 4:1495-1505 (1992) andSpencer et al., Plant Mol. Biol. 24:51-61 (1994).

Any DNA sequence(s), whether from a different species or from the samespecies that is inserted into the genome using transformation isreferred to herein collectively as “transgenes.” In some embodiments ofthe invention, a transformed variant of hybrid cantaloupe BRONCO maycontain at least one transgene but could contain at least 1, 2, 3, 4, 5,6, 7, 8, 9, or 10 transgenes. In another embodiment of the invention, atransformed variant of the another cantaloupe plant used as the donorline may contain at least one transgene but could contain at least 1, 2,3, 4, 5, 6, 7, 8, 9, or 10 transgenes.

Following transformation of cantaloupe target tissues, expression of theabove-described selectable marker genes allows for preferentialselection of transformed cells, tissues and/or plants, usingregeneration and selection methods now well known in the art.

The foregoing methods for transformation would typically be used forproducing a transgenic inbred line or a transgenic hybrid plant. Thetransgenic inbred line could then be crossed, with another(non-transformed or transformed) inbred line, in order to produce a newtransgenic inbred line or plant. Alternatively, a genetic trait whichhas been engineered into a particular cantaloupe plant using theforegoing transformation techniques could be moved into anothercantaloupe plant using traditional backcrossing techniques that are wellknown in the plant breeding arts. For example, a backcrossing approachcould be used to move an engineered trait from a public, non-eliteinbred line into an elite inbred line, or from an inbred line containinga foreign gene in its genome into an inbred line or lines which do notcontain that gene. As used herein, “crossing” can refer to a simple X byY cross, or the process of backcrossing, depending on the context.

iii Selection

For efficient plant transformation, a selection method must be employedsuch that whole plants are regenerated from a single transformed celland every cell of the transformed plant carries the DNA of interest.These methods can employ positive selection, whereby a foreign gene issupplied to a plant cell that allows it to utilize a substrate presentin the medium that it otherwise could not use, such as mannose or xylose(for example, refer U.S. Pat. No. 5,767,378; U.S. Pat. No. 5,994,629).More typically, however, negative selection is used because it is moreefficient, utilizing selective agents such as herbicides or antibioticsthat either kill or inhibit the growth of non-transformed plant cellsand reducing the possibility of chimeras. Resistance genes that areeffective against negative selective agents are provided on theintroduced foreign DNA used for the plant transformation. For example,one of the most popular selective agents used is the antibiotickanamycin, together with the resistance gene neomycin phosphotransferase(nptII), which confers resistance to kanamycin and related antibiotics(see, for example, Messing & Vierra, Gene 19: 259-268 (1982); Bevan etal., Nature 304:184-187 (1983)). However, many different antibiotics andantibiotic resistance genes can be used for transformation purposes(refer U.S. Pat. No. 5,034,322, U.S. Pat. No. 6,174,724 and U.S. Pat.No. 6,255,560). In addition, several herbicides and herbicide resistancegenes have been used for transformation purposes, including the bargene, which confers resistance to the herbicide phosphinothricin (Whiteet al., Nucl Acids Res 18: 1062 (1990), Spencer et al., Theor Appl Genet79: 625-631 (1990), U.S. Pat. No. 4,795,855, U.S. Pat. No. 5,378,824 andU.S. Pat. No. 6,107,549). In addition, the dhfr gene, which confersresistance to the anticancer agent methotrexate, has been used forselection (Bourouis et al., EMBO J. 2(7): 1099-1104 (1983).

Additional selectable marker genes of bacterial origin that conferresistance to antibiotics include gentamycin acetyl transferase,streptomycin phosphotransferase, and aminoglycoside-3′-adenyltransferase, the bleomycin resistance determinant. Hayford et al., PlantPhysiol. 86:1216 (1988), Jones et al., Mol. Gen. Genet., 210:86 (1987),Svab et al., Plant Mol. Biol. 14:197 (1990), Hille et al., Plant Mol.Biol. 7:171 (1986). Other selectable marker genes confer resistance toherbicides such as glyphosate, glufosinate or bromoxynil. Comai et al.,Nature 317:741-744 (1985), Gordon-Kamm et al., Plant Cell 2:603-618(1990) and Stalker et al., Science 242:419-423 (1988).

Other selectable marker genes for plant transformation are not ofbacterial origin. These genes include, for example, mouse dihydrofolatereductase, plant 5-enolpyruvylshikimate-3-phosphate synthase and plantacetolactate synthase. Eichholtz et al., Somatic Cell Mol. Genet. 13:67(1987), Shah et al., Science 233:478 (1986), Charest et al., Plant CellRep. 8:643 (1990).

Another class of marker genes for plant transformation requiresscreening of presumptively transformed plant cells rather than directgenetic selection of transformed cells for resistance to a toxicsubstance such as an antibiotic. These genes are particularly useful toquantify or visualize the spatial pattern of expression of a gene inspecific tissues and are frequently referred to as reporter genesbecause they can be fused to a gene or gene regulatory sequence for theinvestigation of gene expression. Commonly used genes for screeningpresumptively transformed cells include beta-glucuronidase (GUS,beta-galactosidase, luciferase and chloramphenicol acetyltransferase.Jefferson, R. A., Plant Mol. Biol. Rep. 5:387 (1987), Teeri et al., EMBOJ. 8:343 (1989), Koncz et al., Proc. Natl. Acad. Sci U.S.A. 84:131(1987), DeBlock et al., EMBO J. 3:1681 (1984), Valles et al, Plant CellReport 3:3-4 145-148 (1994), Shetty et al., FoodBiotechnology 11:2111-128 (1997)

In vivo methods for visualizing GUS activity that do not requiredestruction of plant tissue are also available. However, these in vivomethods for visualizing GUS activity have not proven useful for recoveryof transformed cells because of low sensitivity, high fluorescentbackgrounds and limitations associated with the use of luciferase genesas selectable markers. A gene encoding Green Fluorescent Protein (GFP)has been utilized as a marker for gene expression in prokaryotic andeukaryotic cells. Chalfie et al., Science 263:802 (1994). GFP andmutants of GFP may be used as screenable markers.

iv Expression Vectors

Genes can be introduced in a site directed fashion using homologousrecombination. Homologous recombination permits site-specificmodifications in endogenous genes and thus inherited or acquiredmutations may be corrected, and/or novel alterations may be engineeredinto the genome. Homologous recombination and site-directed integrationin plants are discussed in, for example, U.S. Pat. Nos. 5,451,513;5,501,967 and 5,527,695.

The expression control elements used to regulate the expression of agiven protein can either be the expression control element that isnormally found associated with the coding sequence (homologousexpression element) or can be a heterologous expression control element.A variety of homologous and heterologous expression control elements areknown in the art and can readily be used to make expression units foruse in the present invention. Transcription initiation regions, forexample, can include any of the various opine initiation regions, suchas octopine, mannopine, nopaline and the like that are found in the Tiplasmids of Agrobacterium tumefaciens. Alternatively, plant viralpromoters can also be used, such as the cauliflower mosaic virus 19S and35S promoters (CaMV 19S and CaMV 35S promoters, respectively) to controlgene expression in a plant (U.S. Pat. Nos. 5,352,605; 5,530,196 and5,858,742 for example). Enhancer sequences derived from the CaMV canalso be utilized (U.S. Pat. Nos. 5,164,316; 5,196,525; 5,322,938;5,530,196; 5,352,605; 5,359,142; and 5,858,742 for example). Lastly,plant promoters such as prolifera promoter, fruit specific promoters,Ap3 promoter, heat shock promoters, seed specific promoters, etc. canalso be used.

Either a gamete-specific promoter, a constitutive promoter (such as theCaMV or Nos promoter), an organ-specific promoter (such as the E8promoter from tomato), or an inducible promoter is typically ligated tothe protein or antisense encoding region using standard techniques knownin the art. The expression unit may be further optimized by employingsupplemental elements such as transcription terminators and/or enhancerelements.

Thus, for expression in plants, the expression units will typicallycontain, in addition to the protein sequence, a plant promoter region, atranscription initiation site and a transcription termination sequence.Unique restriction enzyme sites at the 5′ and 3′ ends of the expressionunit are typically included to allow for easy insertion into apre-existing vector.

In the construction of heterologous promoter/structural gene orantisense combinations, the promoter is preferably positioned about thesame distance from the heterologous transcription start site as it isfrom the transcription start site in its natural setting. As is known inthe art, however, some variation in this distance can be accommodatedwithout loss of promoter function.

In addition to a promoter sequence, the expression cassette can alsocontain a transcription termination region downstream of the structuralgene to provide for efficient termination. The termination region may beobtained from the same gene as the promoter sequence or may be obtainedfrom different genes. If the mRNA encoded by the structural gene is tobe efficiently processed, DNA sequences which direct polyadenylation ofthe RNA are also commonly added to the vector construct. Polyadenylationsequences include, but are not limited to the Agrobacterium octopinesynthase signal (Gielen et al., EMBO J 3:835-846 (1984)) or the nopalinesynthase signal (Depicker et al., Mol. and Appl. Genet. 1:561-573(1982)). The resulting expression unit is ligated into or otherwiseconstructed to be included in a vector that is appropriate for higherplant transformation. One or more expression units may be included inthe same vector. The vector will typically contain a selectable markergene expression unit by which transformed plant cells can be identifiedin culture. Usually, the marker gene will encode resistance to anantibiotic, such as G418, hygromycin, bleomycin, kanamycin, orgentamicin or to an herbicide, such as glyphosate (Round-Up) orglufosinate (BASTA) or atrazine. Replication sequences, of bacterial orviral origin, are generally also included to allow the vector to becloned in a bacterial or phage host; preferably a broad host range forprokaryotic origin of replication is included. A selectable marker forbacteria may also be included to allow selection of bacterial cellsbearing the desired construct. Suitable prokaryotic selectable markersinclude resistance to antibiotics such as ampicillin, kanamycin ortetracycline. Other DNA sequences encoding additional functions may alsobe present in the vector, as is known in the art. For instance, in thecase of Agrobacterium transformations, T-DNA sequences will also beincluded for subsequent transfer to plant chromosomes.

To introduce a desired gene or set of genes by conventional methodsrequires a sexual cross between two lines, and then repeatedback-crossing between hybrid offspring and one of the parents until aplant with the desired characteristics is obtained. This process,however, is restricted to plants that can sexually hybridize, and genesin addition to the desired gene will be transferred.

Recombinant DNA techniques allow plant researchers to circumvent theselimitations by enabling plant geneticists to identify and clone specificgenes for desirable traits, such as improved fatty acid composition, andto introduce these genes into already useful varieties of plants. Oncethe foreign genes have been introduced into a plant, that plant can thenbe used in conventional plant breeding schemes (e.g., pedigree breeding,single-seed-descent breeding schemes, reciprocal recurrent selection) toproduce progeny which also contain the gene of interest.

v Promoters

Genes included in expression vectors must be driven by nucleotidesequence comprising a regulatory element, for example, a promoter.Several types of promoters are now well known in the transformationarts, as are other regulatory elements that can be used alone or incombination with promoters.

As used herein, “promoter” includes reference to a region of DNAupstream from the start of transcription and involved in recognition andbinding of RNA polymerase and other proteins to initiate transcription.A “plant promoter” is a promoter capable of initiating transcription inplant cells. Examples of promoters under developmental control includepromoters that preferentially initiate transcription in certain organs,such as leaves, roots, seeds and tissues such as fibers, xylem vessels,tracheids, or sclerenchyma. Such promoters are referred to as“tissue-preferred”. Promoters which initiate transcription only incertain tissue are referred to as “tissue-specific”. A “cell type”specific promoter primarily drives expression in certain cell types inone or more organs, for example, vascular cells in roots or leaves. An“inducible” promoter is a promoter which is under environmental control.Examples of environmental conditions that may effect transcription byinducible promoters include anaerobic conditions or the presence oflight. Tissue-specific, tissue-preferred, cell type specific, andinducible promoters constitute the class of “non-constitutive”promoters. A “constitutive” promoter is a promoter which is active undermost environmental conditions.

A) Inducible Promoters

An inducible promoter is operably linked to a gene for expression incantaloupe. Optionally, the inducible promoter is operably linked to anucleotide sequence encoding a signal sequence which is operably linkedto a gene for expression in cantaloupe. With an inducible promoter therate of transcription increases in response to an inducing agent.

Any inducible promoter can be used in the instant invention. See Ward etal., Plant Mol. Biol. 22:361-366 (1993). Exemplary inducible promotersinclude, but are not limited to, that from the ACEI system whichresponds to copper (Mett et al., PNAS 90:4567-4571 (1993)); In2 genefrom maize which responds to benzenesulfonamide herbicide safeners (Gatzet al., Mol. Gen. Genetics 243:32-38 (1994)) or Tet repressor from Tn10(Gatz et al., Mol. Gen. Genetics 227:229-237 (1991)). A particularlypreferred inducible promoter is a promoter that responds to an inducingagent to which plants do not normally respond. An exemplary induciblepromoter is the inducible promoter from a steroid hormone gene, thetranscriptional activity of which is induced by a glucocorticosteroidhormone (Schena et al., Proc. Natl. Acad. Sci. U.S.A. 88:0421 (1991)).

B) Constitutive Promoters

A constitutive promoter is operably linked to a gene for expression incantaloupe or the constitutive promoter is operably linked to anucleotide sequence encoding a signal sequence which is operably linkedto a gene for expression in cantaloupe.

Many different constitutive promoters can be utilized in the instantinvention. Exemplary constitutive promoters include, but are not limitedto, the promoters from plant viruses such as the 35S promoter from CaMV(Odell et al., Nature 313:810-812 (1985)) and the promoters from suchgenes as rice actin (McElroy et al., Plant Cell 2:163-171 (1990));ubiquitin (Christensen et al., Plant Mol. Biol. 12:619-632 (1989) andChristensen et al., Plant Mol. Biol. 18:675-689 (1992)); pEMU (Last etal., Theor. Appl. Genet. 81:581-588 (1991)); MAS (Velten et al., EMBO J.3:2723-2730 (1984)) and maize H3 histone (Lepetit et al., Mol. Gen.Genetics 231:276-285 (1992) and Atanassova et al., Plant Journal 2 (3):291-300 (1992)). The ALS promoter, Xba1/Ncol fragment 5′ to the Brassicanapus ALS3 structural gene (or a nucleotide sequence similarity to saidXba1/Ncol fragment), represents a particularly useful constitutivepromoter. See PCT application WO96/30530.C. Tissue-specific orTissue-preferred Promoters

A tissue-specific promoter is operably linked to a gene for expressionin cantaloupe. Optionally, the tissue-specific promoter is operablylinked to a nucleotide sequence encoding a signal sequence which isoperably linked to a gene for expression in cantaloupe. Plantstransformed with a gene of interest operably linked to a tissue-specificpromoter produce the protein product of the transgene exclusively, orpreferentially, in a specific tissue.

Any tissue-specific or tissue-preferred promoter can be utilized in theinstant invention. Exemplary tissue-specific or tissue-preferredpromoters include, but are not limited to, a root-preferred promoter,such as that from the phaseolin gene (Murai et al., Science 23:476-482(1983) and Sengupta-Gopalan et al., Proc. Natl. Acad. Sci. U.S.A.82:3320-3324 (1985)); a leaf-specific and light-induced promoter such asthat from cab or rubisco (Simpson et al., EMBO J. 4(11):2723-2729 (1985)and Timko et al., Nature 318:579-582 (1985)); an anther-specificpromoter such as that from LAT52 (Twell et al., Mol. Gen. Genetics217:240-245 (1989)); a pollen-specific promoter such as that from Zm13or a microspore-preferred promoter such as that from apg (Twell et al.,Sex. Plant Reprod. 6:217-224 (1993)).

Signal Sequences for Targeting Proteins to Subcellular Compartments

Transport of protein produced by transgenes to a subcellular compartmentsuch as the chloroplast, vacuole, peroxisome, glyoxysome, cell wall ormitochondrion or for secretion into the apoplast, is accomplished bymeans of operably linking the nucleotide sequence encoding a signalsequence to the 5′ and/or 3′ region of a gene encoding the protein ofinterest. Targeting sequences at the 5′ and/or 3′ end of the structuralgene may determine, during protein synthesis and processing, where theencoded protein is ultimately compartmentalized.

The presence of a signal sequence directs a polypeptide to either anintracellular organelle or subcellular compartment or for secretion tothe apoplast. Many signal sequences are known in the art. See, forexample Becker et al., Plant Mol. Biol. 20:49 (1992), Knox, C., et al.,Plant Mol. Biol. 9:3-17 (1987), Lerner et al., Plant Physiol. 91:124-129(1989), Fontes et al., Plant Cell 3:483-496 (1991), Matsuoka et al.,Proc. Natl. Acad. Sci. 88:834 (1991), Gould et al., J. Cell. Biol.108:1657 (1989), Creissen et al., Plant J. 2:129 (1991), Kalderon, etal., Cell 39:499-509 (1984), Stiefel, et al., Plant Cell 2:785-793(1990).

Foreign Protein Genes and Agronomic Genes

With transgenic plants according to the present invention, a foreignprotein can be produced in commercial quantities. Thus, techniques forthe selection and propagation of transformed plants, which are wellunderstood in the art, yield a plurality of transgenic plants which areharvested in a conventional manner, and a foreign protein then can beextracted from a tissue of interest or from total biomass. Proteinextraction from plant biomass can be accomplished by known methods whichare discussed, for example, by Heney and Orr, Anal. Biochem. 114:92-6(1981).

According to a one embodiment, the transgenic plant provided forcommercial production of foreign protein is cantaloupe plant. In anotherpreferred embodiment, the biomass of interest is seed. For therelatively small number of transgenic plants that show higher levels ofexpression, a genetic map can be generated, primarily via conventionalRFLP, PCR and SSR analysis, which identifies the approximate chromosomallocation of the integrated DNA molecule. For exemplary methodologies inthis regard, see Glick and Thompson, Methods in Plant Molecular Biologyand Biotechnology, Glick and Thompson Eds., CRC Press, Boca Raton269:284 (1993). Map information concerning chromosomal location isuseful for proprietary protection of a subject transgenic plant. Ifunauthorized propagation is undertaken and crosses made with othergermplasm, the map of the integration region can be compared to similarmaps for suspect plants, to determine if the latter have a commonparentage with the subject plant. Map comparisons would involvehybridizations, RFLP, PCR, SSR and sequencing, all of which areconventional techniques.

Likewise, by means of the present invention, agronomic genes can beexpressed in transformed plants. More particularly, plants can begenetically engineered to express various phenotypes of agronomicinterest. Exemplary genes implicated in this regard include, but are notlimited to, those categorized below:

Genes that Confer Resistance to Pests or Disease and that Encode:

A. Plant disease resistance genes. Plant defenses are often activated byspecific interaction between the product of a disease resistance gene(R) in the plant and the product of a corresponding avirulence (Avr)gene in the pathogen. A plant variety can be transformed with one ormore cloned resistance gene to engineer plants that are resistant tospecific pathogen strains. See, for example Jones et al., Science266:789 (1994) (cloning of the tomato Cf-9 gene for resistance toCladosporium fulvum); Martin et al., Science 262:1432 (1993) (tomato Ptogene for resistance to Pseudomonas syringae pv. tomato encodes a proteinkinase); Mindrinos et al., Cell 78:1089 (1994) (Arabidopsis RSP2 genefor resistance to Pseudomonas syringae).

B. A Bacillus thuringiensis protein, a derivative thereof or a syntheticpolypeptide modeled thereon. See, for example, Geiser et al., Gene48:109 (1986), who disclose the cloning and nucleotide sequence of a Btalpha-endotoxin gene. Moreover, DNA molecules encoding alpha-endotoxingenes can be purchased from American Type Culture Collection, Manassas,Va., for example, under ATCC Accession Nos. 40098, 67136, 31995 and31998.

C. A lectin. See, for example, the disclosure by Van Damme et al., PlantMolec. Biol. 24:25 (1994), who disclose the nucleotide sequences ofseveral Clivia miniata mannose-binding lectin genes.

D. A vitamin-binding protein such as avidin. See PCT applicationUS93/06487. The application teaches the use of avidin and avidinhomologues as larvicides against insect pests.

E. An enzyme inhibitor, for example, a protease or proteinase inhibitoror an amylase inhibitor. See, for example, Abe et al., J. Biol. Chem.262:16793 (1987) (nucleotide sequence of rice cysteine proteinaseinhibitor), Huub et al., Plant Molec. Biol. 21:985 (1993) (nucleotidesequence of cDNA encoding tobacco proteinase inhibitor I), Sumitani etal., Biosci. Biotech. Biochem. 57:1243 (1993) (nucleotide sequence ofStreptomyces nitrosporeus alpha-amylase inhibitor).

F. An insect-specific hormone or pheromone such as an ecdysteroid andjuvenile hormone, a variant thereof, a mimetic based thereon, or anantagonist or agonist thereof. See, for example, the disclosure byHammock et al., Nature 344:458 (1990), of baculovirus expression ofcloned juvenile hormone esterase, an inactivator of juvenile hormone.

G. An insect-specific peptide or neuropeptide which, upon expression,disrupts the physiology of the affected pest. Pratt et al., Biochem.Biophys. Res. Comm. 163:1243 (1989) (an allostatin is identified inDiploptera puntata). See also U.S. Pat. No. 5,266,317 to Tomalski etal., who disclose genes encoding insect-specific, paralytic neurotoxins.

H. An insect-specific venom produced in nature by a snake, a wasp, etc.For example, see Pang et al., Gene 116:165 (1992), for disclosure ofheterologous expression in plants of a gene coding for a scorpioninsectotoxic peptide.

I. An enzyme responsible for a hyper-accumulation of a monoterpene, asesquiterpene, a steroid, a hydroxamic acid, a phenylpropanoidderivative or another non-protein molecule with insecticidal activity.

J. An enzyme involved in the modification, including thepost-translational modification, of a biologically active molecule; forexample, a glycolytic enzyme, a proteolytic enzyme, a lipolytic enzyme,a nuclease, a cyclase, a transaminase, an esterase, a hydrolase, aphosphatase, a kinase, a phosphorylase, a polymerase, an elastase, achitinase and a glucanase, whether natural or synthetic. See PCTapplication WO 93/02197 in the name of Scott et al., which discloses thenucleotide sequence of a callase gene. DNA molecules which containchitinase-encoding sequences can be obtained, for example, from the ATCCunder Accession Nos. 39637 and 67152. See also Kramer et al., InsectBiochem. Molec. Biol. 23:691 (1993), who teach the nucleotide sequenceof a cDNA encoding tobacco hornworm chitinase, and Kawalleck et al.,Plant Molec. Biol. 21:673 (1993), who provide the nucleotide sequence ofthe parsley ubi4-2 polyubiquitin gene.

K. A molecule that stimulates signal transduction. For example, see thedisclosure by Botella et al., Plant Molec. Biol. 24:757 (1994), ofnucleotide sequences for mung bean calmodulin cDNA clones, and Griess etal., Plant Physiol. 104:1467 (1994), who provide the nucleotide sequenceof a maize calmodulin cDNA clone.

L. A hydrophobic moment peptide. See PCT application WO95/16776(disclosure of peptide derivatives of Tachyplesin which inhibit fungalplant pathogens) and PCT application WO95/18855 (teaches syntheticantimicrobial peptides that confer disease resistance).

M. A membrane permease, a channel former or a channel blocker. Forexample, see the disclosure of Jaynes et al., Plant Sci. 89:43 (1993),of heterologous expression of a cecropin-beta, lytic peptide analog torender transgenic tobacco plants resistant to Pseudomonas solanacearum.

N. A viral-invasive protein or a complex toxin derived therefrom. Forexample, the accumulation of viral coat proteins in transformed plantcells imparts resistance to viral infection and/or disease developmenteffected by the virus from which the coat protein gene is derived, aswell as by related viruses. See Beachy et al., Ann. Rev. Phytopathol.28:451 (1990). Coat protein-mediated resistance has been conferred upontransformed plants against alfalfa mosaic virus, cucumber mosaic virus,tobacco streak virus, potato virus X, potato virus Y, tobacco etchvirus, tobacco rattle virus and tobacco mosaic virus. Id.

O. An insect-specific antibody or an immunotoxin derived therefrom.Thus, an antibody targeted to a critical metabolic function in theinsect gut would inactivate an affected enzyme, killing the insect

P. A virus-specific antibody. See, for example, Tavladoraki et al.,Nature 366:469 (1993), who show that transgenic plants expressingrecombinant antibody genes are protected from virus attack.

Q. A developmental-arrestive protein produced in nature by a pathogen ora parasite. Thus, fungal endo-alpha-1, 4-D-polygalacturonases facilitatefungal colonization and plant nutrient release by solubilizing plantcell wall homo-alpha-1, 4-D-galacturonase. See Lamb et al.,BioTechnology 10:1436 (1992). The cloning and characterization of a genewhich encodes a bean endopolygalacturonase-inhibiting protein isdescribed by Toubart et al., Plant J. 2:367 (1992).

R. A developmental-arrestive protein produced in nature by a plant. Forexample, Logemann et al., BioTechnology 10:305 (1992), have shown thattransgenic plants expressing the barley ribosome-inactivating gene havean increased resistance to fungal disease.

Genes that Confer Resistance to an Herbicide, for Example:

A. An herbicide that inhibits the growing point or meristem, such as animidazolinone or a sulfonylurea. Exemplary genes in this category codefor mutant ALS and AHAS enzyme as described, for example, by Lee et al.,EMBO J. 7:1241 (1988), and Miki et al., Theor. Appl. Genet. 80:449(1990), respectively.

B. Glyphosate (resistance conferred by mutant 5-enolpyruvlshikimate-3phosphate synthase (EPSP) and aroA genes, respectively) and otherphosphono compounds such as glufosinate (phosphinothricin acetyltransferase (PAT) and Streptomyces hygroscopicus PAT, bar, genes), andpyridinoxy or phenoxy propionic acids and cyclohexones (ACCaseinhibitor-encoding genes). See, for example, U.S. Pat. No. 4,940,835 toShah, et al., which discloses the nucleotide sequence of a form of EPSPwhich can confer glyphosate resistance. A DNA molecule encoding a mutantaroA gene can be obtained under ATCC accession number 39256, and thenucleotide sequence of the mutant gene is disclosed in U.S. Pat. No.4,769,061 to Comai. European patent application No. 0 333 033 to Kumadaet al., and U.S. Pat. No. 4,975,374 to Goodman et al., disclosenucleotide sequences of glutamine synthatase genes which conferresistance to herbicides such as L-phosphinothricin. The nucleotidesequence of a PAT gene is provided in European application No. 0 242 246to Leemans et al. DeGreef et al., BioTechnology 7:61 (1989), describethe production of transgenic plants that express chimeric bar genescoding for PAT activity. Exemplary of genes conferring resistance tophenoxy propionic acids and cyclohexones, such as sethoxydim andhaloxyfop are the Acc1-S1, Acc1-S2 and Acc1-S3 genes described byMarshall et al., Theor. Appl. Genet. 83:435 (1992).

C. An herbicide that inhibits photosynthesis, such as a triazine (psbAand gs+ genes) or a benzonitrile (nitrilase gene). Przibilla et al.,Plant Cell 3:169 (1991), describe the transformation of Chlamydomonaswith plasmids encoding mutant psbA genes. Nucleotide sequences fornitrilase genes are disclosed in U.S. Pat. No. 4,810,648 to Stalker, andDNA molecules containing these genes are available under ATCC AccessionNos. 53435, 67441, and 67442. Cloning and expression of DNA coding for aglutathione S-transferase is described by Hayes et al., Biochem. J.285:173 (1992).

Genes that Confer or Contribute to a Value-Added Trait, Such as:

A. Increased sweetness and flavor of the fruit by introduction of a geneencoding sweet-tasting proteins such as monellin (Penarrubia et al.,Biotechnology. 1992, 10: 5, 561-564) or thaumatin (Bartoszewski et al,Plant Breeding 122, 347-351 (2003)).

B. Reduced ethylene biosynthesis to control ripening by introduction ofan antisense construct of the ACC oxidase into Cucumis melo. Forexample, see Ayub et al, Nature Biotechnology 14: 862 (1996)

C. Delayed senescence and improved ripening control by transferring agene or acting on the transcription of a gene involved in plantsenescence. See Wang et al. in Plant Mol. Bio. 52:1223-1235 (2003) onthe role of the deoxyhypusine synthase in the senescence. See also U.S.Pat. No. 6,538,182 issued Mar. 25, 2003.

D. Improved salt tolerance by transforming Cucumis melo plant with HAL1, a yeast regulatory gene involved in stress tolerance, as shown inSerrano et al., Scientia Horticulturae. 1999, 78: 1/4, 261-269 or inBordas et al., Transgenic Research. 1997, 6: 1, 41-50.

E. Obtained male sterile plants, especially useful in hybrid melonproduction, by introduction of a gene encoding a tobacco PR Glucanase asdescribed in tomato (WO9738116) but that can also be used in melon.

Tissue Culture

As it is well known in the art, tissue culture of cantaloupe can be usedfor the in vitro regeneration of cantaloupe plants. Tissues cultures ofvarious tissues of cantaloupe and regeneration of plants therefrom arewell known and published. By way of example, a tissue culture comprisingorgans has been used to produce regenerated plants as described in DirksR., et al. Plant Cell Report 7: 8 626-627 (1989); Homma, Y., et al.Japan J. Breeding. 41:543-551 (1991). Yoshioka, K., et al. Japan J.Breeding. 42:277-285 (1992); Debeaujon, I., et al. Plant Cell Report12:37-40 (1992); Debeaujon, I., et al. Plant Cell Tissue Organ Culture34:91-100 (1993); Fang, G. W., et al. Molecular Plant—MicrobeInteractions 6:358-367 (1993); Valles, M. P., et al. Plant Cell Report13:145-148 (1994); Ezura, H., et al. Plant Cell Report 14:107-111(1994); Kathal, R., et al. Plant Science 96:137-142 (1994); Adelberg, J.W., et al. Hortscience 29:689-692 (1994). More precisely, in the case ofthe melon (C. melo), regeneration through organogenesis has beendescribed either directly on cotyledons placed in culture (Dirks, R. etal., Plant Cell Report, 7:626-627 (1989)), or through the intermediaryof calli derived from cotyledons (Mackay, W. et al., Cucurbit GeneticsCooperative, 11:33-34 (1988), Orts, M. et al., Hort. Science, 22:666(1987)), hypocotyls (Abak, K. et al., Cucurbit Genetics CooperativeReport, 3:27-29 (1980), Kathal, R. et al., J. Plant Physiol., 126:59-62(1986)) or leaves (Kathal, R. et al., Plant Cell Report, 7:449-451(1988)). The production of melon plants derived from somatic embryos hasalso been reported, Oridate, T. et al., Japan J. Breeding, 36:424-428(1986), Branchard, M. et al., C.R. Acad. Sci. Paris, 307, Serie111:777-780 (1988). Also, De Both et al. in U.S. Pat. No. 6,198,022teach how to regenerate plants having a normal phenotype fromcotyledons. It is clear from the literature that the state of the art issuch that these methods of obtaining plants are routinely used and havea very high rate of success. Thus, another aspect of this invention isto provide cells which upon growth and differentiation producecantaloupe plants having the physiological and morphologicalcharacteristics of hybrid cantaloupe plant BRONCO.

As used herein, the term “tissue culture” indicates a compositioncomprising isolated cells of the same or a different type or acollection of such cells organized into parts of a plant. Exemplarytypes of tissue cultures are protoplasts, calli, plant clumps, and plantcells that can generate tissue culture that are intact in plants orparts of plants, such as embryos, pollen, flowers, seeds, leaves, stems,roots, root tips, anthers, pistils, meristematic cells, axillary buds,ovaries, seed coat, endosperm, hypocotyls, cotyledons and the like.Means for preparing and maintaining plant tissue culture are well knownin the art. By way of example, a tissue culture comprising organs hasbeen used to produce regenerated plants. U.S. Pat. Nos. 5,959,185,5,973,234, and 5,977,445 describe certain techniques, the disclosures ofwhich are incorporated herein by reference.

EXAMPLES

The foregoing examples of the related art and limitations relatedtherewith are intended to be illustrative and not exclusive. Otherlimitations of the related art will become apparent to those of skill inthe art upon a reading of the specification.

Example 1 Development of New BRONCO Cantaloupe Variety

Hybrid cantaloupe plant BRONCO has superior characteristics and has beendeveloped through the crossing of cantaloupe inbred line M056WN withcantaloupe inbred line I407.

Inbred cantaloupe M056WN is a selection from a family of double haploidlines created by the development of haploid individuals from an F1 line.Both parents of the F1 line were elite proprietary lines containingdifferent but complimentary traits. Parent A is a large fruited, earlymaturing, monoecious, fully netted western shipper inbred withresistance to Powdery mildew race 2 (US). Parent A has medium firm fleshwith medium sugar content and medium orange flesh color. Parent B is asmall fruited, later maturing, andromonoecious, heavily netted westernshipper inbred with resistance to Fusarium oxysporum spp. Melonis race 0and 2. Parent B has extremely firm flesh with high sugar content anddark orange flesh color.

The F1 between parent A and parent B was made in Chile in the firstyear. The F1 was planted in the summer of the first year in Davis,Calif. and was pollinated with irradiated pollen to induce thedevelopment of haploid embryos. Haploid embryos were harvested inSeptember of first year, placed on callus media, and then doubled inorder to obtain double haploid plantlets. These double haploid plantswere grown in the greenhouse in Davis in the spring of second year andwere self-pollinated to obtain seed of each DH (double haploid) event.22 unique DH lines were successfully recovered and were planted in theopen field at Davis in for initial evaluation. M056WN was one of the DHevents from this group. In third year, M056WN was sent to Chile and aset of hybrids was developed using this as a new parent. One of thesehybrids was to become the variety Bronco. Concurrently and continuingthrough year four, M056WN was evaluated in multiple locations includingDavis field and Immokalee Fla. It was also screened for resistance toPowdery mildew race 2 (US) and Fusarium oxysporum spp. Melonis race 0and 2. After three years of evaluation M056WN was determined to be anexceptional breeding line combining many desirable traits from Parents Aand B. It is a medium to large size, fully netted western shipperinbred, andromonecious with very dark orange flesh color and firm flesh.It is mid early in maturity and has a strong plant that is broadlyadaptable. It is resistant to Fusarium oxysporum spp. Melonis race 0 and2 and has intermediate resistance (IR) to Powdery mildew race 2 (US).This line also expresses high level of sugar under many differentconditions. It has proven to have excellent combining ability and verybroad utilization.

Inbred I407 was developed by self-pollinating a very successfulcommercial variety. The F1 was self-pollinated in the first year and theline development activity (F2-F9) was conducted at both Davis, Calif.and Ruskin, Fla. It was developed using a pedigree breeding method. Itwas selected for excellent fruit size and shape, medium orange fleshcolor and a very small seed cavity. It is mid early in maturity, with afine, medium closed net. The breeding and pathology screening work onI407 was completed in year five at which time a number of experimentalhybrids were made with I407. It has intermediate resistance to Powderymildew race 2 (US) and is tolerant to sulfur application.

Hybrid cantaloupe plant Bronco is similar to hybrid cantaloupe plant OroRico. While similar to hybrid cantaloupe plant Oro Rico, there aresignificant differences including: Bronco matures at least 4 daysearlier than Oro Rico. Bronco has a more concentrated harvest period,with all marketable fruit maturing within a 10 day period under summerconditions in Davis Calif. Oro Rico has a more extended harvest period,with all marketable fruit maturing within a 16 day period. Fruit weightfor Bronco is approximately 29% larger than Oro Rico. Under postharvestconditions, 11 days at 5 degrees C., the external firmness isapproximately 70% firmer than Oro Rico. While both varieties have goodnetting, Bronco's netting has a more coarse texture based on trial dataat many locations. Data across at least 5 trial locations indicate thatBronco has a soluble solids (brix or sugar level) 1.5 degrees higherthan Oro Rico.

Some of the criteria used to select the hybrids as well as their inbredparent lines in various generations include Vine strength, concentrationof fruit set, yield, netting density and type, fruit shape, fruit size,maturity, flesh color, flesh firmness, cavity size, sex type, brix,immature skin color.

The hybrid cantaloupe plant BRONCO has shown uniformity and stabilityfor the traits, within the limits of environmental influence for thetraits as described in the following Variety Descriptive Information. Novariant traits have been observed or are expected for agronomicalimportant traits in cantaloupe hybrid BRONCO.

Hybrid cantaloupe plant BRONCO has the following morphologic and othercharacteristics, as compared to Oro Rico (based primarily on datacollected in California, all experiments done under the directsupervision of the applicant).

TABLE 1 Variety Description Information BRONCO Variety Plant TypeCucumis melo var. reticulatus Region Where Western U.S.A. Developed Areaof Best Central California, Arizona Adaptation for U.S.A. Maturity 80days (4 days earlier than Oro Rico variety) Leaf Morphology (AverageValues From Mature Blade of Third Leaf) Shape Slightly lobed Length 101mm Width 143 mm Surface Pubescent Fruit Morphology (Average Values atEdible Maturity) Length 17 cm Diameter 16 cm Weight 2130 gm Shape Round,slightly oval Surface Full netted, no vein tracts Blossom Scar Medium,slightly conspicuous Ribs Not present Number of Ribs per N/A fruit RibWidth at Medial N/A Sutures N/A Shipping Quality Excellent Fruit AbciseAt maturity Long Shelf life No Rind Net (At Edible Maturity)Distribution Complete and closed Coarseness Medium course Rind Color(Average Values at Edible Maturity): Primary Color Green Net Color GrayMottling Color Orange Furrow (Suture) N/A Color Flesh Morphology(Average Values at Edible Maturity) Color Near Cavity Deep orange Colorin Center Deep orange Color near rind Orange Refractometer- 12.8%(Compared to 11.9% SSC for Oro Rico) Soluble Solids Aroma Fruity andslightly musky Flavor Moderate cantaloupe flavor Seed Cavity (AverageValues at Edible Maturity) Length 60 mm Width 60 mm Shape in Cross-Triangulate Section Disease Resistance - Rating (1 = susceptible - 5 =resistant) Bacterial Wilt 1 Powdery Mildew 3 (Podosphaera xanthii andErysiphe cichoracearum) Watermelon Mosaic 1 Anthracnose 1 Root Rot 1Verticillum Wilt 3 Cucumber Mosaic 1 Fusarium Wilt 5 (to U.S. races 0.2)(fusarium oxysporum 1 (to U.S. race 1) spp. Melonis) Melon Rust 1

Example 2 Comparison of New BRONCO Cantaloupe with Check Variety

In the tables that follow, the traits and characteristics of hybridcantaloupe BRONCO are given compared to another hybrid. The datacollected are presented for key characteristics and traits. Hybridcantaloupe BRONCO was tested at numerous locations, with two or threereplications per location. Information about the hybrid, as compared toseveral check hybrid is presented.

Table 2 below shows the characteristics of hybrid cantaloupe BRONCO ascompared to hybrid Oro Rico. Column 1 (Variety ID) identifies the plant,column 2 (Weight) describes the fruit weight which is measured inkilograms, columns 3 (ExtFirm_P1) and 4 (ExtFirm_P2) show the average ofexterior fruit firmness measured in lbs/square inch resistance (2readings (P1 and P2) per fruit after netting being removed), column 5(ExtColor) shows the exterior color, i.e. a subjective rating on a 1-4scale where 1 is green appearance and 4 is ripe appearance, column 6(MOTTLE) shows the mottle, i.e. a subjective rating on a 1-4 scale forthe presence of a mottled ripening appearance where 1 is no mottle and 4is excessive mottle, column 7 (SurfaceMold) shows the average of surfacemold, i.e. a subjective rating on a 1-4 scale for the presence of moldon the fruit surface where 1 is no mold and 4 is heavy mold.

TABLE 2 VarietyID WEIGHT ExtFirm_P1 ExtFirm_P2 ExtColor MOTTLESurfaceMold ORO 1.41 6.98 7.00 3.14 1.53 1.03 RICO BRONCO 1.82 13.3611.74 3.21 1.21 1.21

Table 3 below shows the characteristics of hybrid cantaloupe BRONCO ascompared to hybrid Oro Rico. Column 1 (VarietyID) identifies the plant,column 2 (IntColor) describes the interior color i.e. a subjectiverating on a 1-4 scale for the color where 1 is very pale orange and 4 isvery dark orange, column 3 (IntFirm_P1) and 4 (IntFirm_P2) show theinterior fruit firmness measured in lbs/square inch resistance (2readings (P1 and P2) per fruit at 90 degrees and 270 degrees in centerof flesh between rind and cavity, column 5 (Avg of BRIX) shows the brix,i.e. the sugar content measured in degrees brix.

TABLE 3 VarietyID IntColor IntFirm_P1 IntFirm_P2 Avg Of BRIX ORO 2.892.38 2.44 9.91 RICO BRONCO 3.11 3.07 3.06 9.57

Table 4 below shows the characteristics of hybrid cantaloupe BRONCO ascompared to hybrid Oro Rico. Column 1 (Variety) identifies the plant,column 2 (Vine) describes the vine as an indication of the strength ofthe vine, where “str” is strong and “avg” is average, column 3 (Yield)describes the yield as a subjective determination of the yield potentialwhere “lo” is lower yielding, “avg” is average yielding and “hi” is highyielding, column 4 (Concentration of yield) shows concentration of yieldon the plant where “con” is concentrated set and “sem” is semiconcentrated set, column 5 (Maturity) shows the relative maturity ofeach variety based on ripening fruit where “me” is mid early, “m” ismain maturity and “l” is late maturity, column 6 (Net Spacing) shows thenet spacing, i.e. the relative degree of openness of netting on fruitwhere “med” is medium net spacing and “cl” is closed net spacing, column7 (Net Type) shows the net type, i.e. the relative thickness and widthof fruit netting where “med” is medium texture net and “med cs” ismedium coarse netting, column 8 (Fruit Shape) shows the shape of thefruit from “rnd” which is round to “rslov” which is round to slightoval, column 9 (Fruit Size) shows the fruit size which is represented bythe number of fruit that will fit into a standardized box where 9 equals9 fruits per box a box, 12 equals 12 fruit per box etc., column 10(Brix) shows the brix, i.e. the sugar content measured in degrees brix.

TABLE 4 Concentration Net Net Fruit Fruit Variety Vine Yield of yieldMaturity Spacing Type Shape Size Brix BRONCO str avg sem me med med rnd9-12 14.4 ORO avg hi sem m med med rnd 12-15  11 RICO BRONCO avg lo seml cl Medcs rslov 9 13.8 ORO str hi sem me med med rnd 9-12 11.4 RICOBRONCO avg hi sem me med Medcs ov 9 12.5 ORO str hi sem m med med mov9-12 12 RICO BRONCO str hi sem m med med rnd 9-12 14 ORO avg hi con lmed med rnd 12-15  12 RICODeposit Information

A deposit of the cantaloupe seed of this invention is maintained byHM.CLAUSE, Inc. Davis Research Station, 9241 Mace Boulevard, Davis,Calif. 95616. In addition, a sample of the hybrid cantaloupe seed ofthis invention has been deposited with the National Collections ofIndustrial, Food and Marine Bacteria (NCIMB), 23 St Machar Drive,Aberdeen, Scotland, AB24 3RY, United Kingdom.

To satisfy the enablement requirements of 35 U.S.C. 112, and to certifythat the deposit of the isolated strain of the present invention meetsthe criteria set forth in 37 CFR 1.801-1.809, Applicants hereby make thefollowing statements regarding the deposited hybrid cantaloupe BRONCO(deposited as NCIMB Accession No. 42565):

-   1. During the pendency of this application, access to the invention    will be afforded to the Commissioner upon request;-   2. Upon granting of the patent the strain will be available to the    public under conditions specified in 37 CFR 1.808;-   3. The deposit will be maintained in a public repository for a    period of 30 years or 5 years after the last request or for the    effective life of the patent, whichever is longer; and-   4. The deposit will be replaced if it should ever become    unavailable.    Access to this deposit will be available during the pendency of this    application to persons determined by the Commissioner of Patents and    Trademarks to be entitled thereto under 37 C.F.R. §1.14 and 35    U.S.C. §122. Upon allowance of any claims in this application, all    restrictions on the availability to the public of the variety will    be irrevocably removed by affording access to a deposit of at least    2,500 seeds of the same variety with the NCIMB.

INCORPORATION BY REFERENCE

All references, articles, publications, patents, patent publications,and patent applications cited herein are incorporated by reference intheir entireties for all purposes.

However, mention of any reference, article, publication, patent, patentpublication, and patent application cited herein is not, and should notbe taken as, an acknowledgment or any form of suggestion that theyconstitute valid prior art or form part of the common general knowledgein any country in the world.

What is claimed is:
 1. A seed of hybrid cantaloupe designated BRONCO,wherein a representative sample of seed of said hybrid was depositedunder NCIMB No.
 42565. 2. A cantaloupe plant, or a part thereof,produced by growing the seed of claim
 1. 3. The cantaloupe part of claim2, wherein the cantaloupe part is selected from the group consisting of:a leaf, a flower, a fruit, an ovule, pollen, a cell a rootstock, and ascion.
 4. A cantaloupe plant, or a part thereof, having all of thecharacteristics of hybrid BRONCO listed in Table
 1. 5. A cantaloupeplant, or a part thereof, having all of the physiological andmorphological characteristics of hybrid BRONCO, wherein a representativesample of seed of said hybrid was deposited under NCIMB No.
 42565. 6. Atissue culture of regenerable cells produced from the plant or plantpart of claim 2, wherein cells of the tissue culture are produced from aplant part selected from the group consisting of protoplasts, embryos,meristematic cells, callus, pollen, ovules, flowers, seeds, leaves,roots, root tips, anthers, stems, petioles, fruits, cotyledons andhypocotyls.
 7. A protoplast produced from the tissue culture of claim 6.8. A cantaloupe plant regenerated from the tissue culture of claim 6,said plant having the characteristics of hybrid BRONCO, wherein arepresentative sample of seed of said hybrid was deposited under NCIMBNo.
 42565. 9. A cantaloupe fruit produced from the plant of claim
 2. 10.A method for producing a cantaloupe fruit comprising a) growing thecantaloupe plant of claim 2 to produce a cantaloupe fruit, and b)harvesting said cantaloupe fruit.
 11. A cantaloupe fruit produced by themethod of claim
 10. 12. A method for producing a cantaloupe seedcomprising crossing a first parent cantaloupe plant with a second parentcantaloupe plant and harvesting the resultant cantaloupe seed, whereinsaid first parent cantaloupe plant and/or second parent cantaloupe plantis the cantaloupe plant of claim
 2. 13. A method for producing acantaloupe seed comprising self-pollinating the cantaloupe plant ofclaim 2 and harvesting the resultant cantaloupe seed.
 14. A method ofvegetatively propagating the cantaloupe plant of claim 2, said methodcomprising a) collecting part of a plant of claim 2 and b) regeneratinga plant from said part.
 15. The method of claim 14 further comprisingharvesting a fruit from said plant.
 16. The plant obtained from themethod of claim
 14. 17. The fruit obtained from the method of claim 15.18. A method of producing a cantaloupe plant derived from the hybridvariety BRONCO, the method comprising the steps of: (a) self-pollinatingthe plant of claim 2 at least once to produce a progeny plant; (b)crossing the progeny plant of step (a) with itself or a secondcantaloupe plant to produce a seed; (c) growing a progeny plant of asubsequent generation from the seed produced in step (b) and crossingthe progeny plant of a subsequent generation with itself or a secondcantaloupe plant to produce a cantaloupe plant derived from the hybridcantaloupe variety BRONCO.
 19. The method of claim 18 further comprisingthe step of: (d) repeating steps b) or c) for at least 1 more generationto produce a cantaloupe plant derived from the hybrid cantaloupe varietyBRONCO.
 20. A method of producing a cantaloupe plant derived from thehybrid variety BRONCO, the method comprising the steps of: (a) crossingthe plant of claim 2 with a second cantaloupe plant to produce a progenyplant; (b) crossing the progeny plant of step (a) with itself or asecond cantaloupe plant to produce a seed; (c) growing a progeny plantof a subsequent generation from the seed produced in step (b) ; (d)crossing the progeny plant of a subsequent generation of step (c) withitself or a second cantaloupe plant to produce a cantaloupe plantderived from the hybrid cantaloupe variety BRONCO.
 21. The method ofclaim 20 further comprising the step of: (d) repeating step b) or c) forat least 1 more generation to produce a cantaloupe plant derived fromthe hybrid cantaloupe variety BRONCO.
 22. A method for producing atransgenic cantaloupe plant, the method comprising crossing a firstcantaloupe plant of claim 2 with a second cantaloupe plant containing atransgene, wherein the transgene of said second cantaloupe plant isintegrated into the genome of the cantaloupe plant progeny resultingfrom said cross, and wherein the transgene confers said cantaloupe plantprogeny with at least one trait selected from the group consisting ofmale sterility, male fertility, herbicide resistance, insect resistance,disease resistance, increased sweetness, increased sugar content,increased flavor, improved ripening control, and improved salttolerance.
 23. A method for producing a transgenic cantaloupe plant, themethod comprising transforming at least one transgene into a hybridcantaloupe BRONCO plant, or a plant part or a plant cell thereof orparental line used for producing the hybrid cantaloupe plant BRONCO, asample seed of said hybrid having been deposited under NCIMB AccessionNo. 42565, thereby producing a transgenic cantaloupe plant.
 24. A plantproduced by the method of claim
 23. 25. The plant of claim 2, whereinsaid plant further comprises a transgene.
 26. The plant of claim 25wherein the transgene confers said plant with a trait selected from thegroup consisting of male sterility, male fertility, herbicideresistance, insect resistance, disease resistance, increased sweetness,increased sugar content, increased flavor, improved ripening control,and improved salt tolerance.